Tag: horticulture

Polyphenols present in a wide range of plant-based foods have received great interest owing to their antioxidant capacity and potential protective effect in reducing cardiovascular disease risk through improvement in vascular function and modulation of inflammation.The interpretation of the influence of polyphenols on cardiovascular health in dietary intervention studies can be complicated due to dynamic bio-availability during the processes of absorption, metabolism, distribution, and excretion. Generally, the absorption of dietary polyphenols is widely dependent on the type and structure of the compound and is often slow and largely incomplete in the small intestine. Therefore, significant quantities of polyphenols are retained in the colon. In addition, the non-absorbed polyphenols are subjected to bio-transformation via the activity of enzymes from various microbial groups . Consequently, the micro-biota-derived metabolites of polyphenols are better absorbed in the gut,which then become an important factor in the health effect of polyphenol-containing foods. Important plant polyphenols and their microbial derivatives are listed in ref . Many of the studies that assess bio-availability and effects of polyphenols have evaluated the balance between the enterohepatic circulating levels, residence time in plasma, and urine excretion rate of the parent phenolic compounds and their microbial-derived metabolites using metabolomic techniques. Importantly, although endogenous enzyme and transporter activities in the small intestine as well as transformation of polyphenols are subject to a wide inter individual variability,nft channel the functional capability of the gut micro-biota is important to partially explain the variation of bio-availability among the population.

Assessing the properties of a single dietary constituent from the polyphenol family alone without dietary fiber is difficult due to the complex dietary food matrix present in a flavonoid-rich diet. For example, regular consumption of apples increased the numbers of fecal bifidobacteria and decreased the C. perf ringens count.Similarly, the concomitant dietary presence of apple polyphenols and FOS increased SCFA production.In contrast, compared to consumption of an inulin-containing diet alone, including a grapefruit flavonoidrich extract decreased both production of SCFAs and the bifidobacteria population.Furthermore, a randomized crossover intervention study in which subjects consumed high levels of cruciferous vegetables for 14 days revealed an alteration of the fecal microbial community profile compared with a low phytochemical, low-fiber diet, including a higher abundance of Eubacterium hallii, Phascolarctobacterium faecium, Burkholderiales spp., Alistipes putredinis and Eggerthella spp.The observed changes could also be explained by the increase in dietary fiber that is enriched in cruciferous vegetables.200 Overall, the direct effects of fiber blur the ability to judge the specific effects of individual dietary ingredients on gut micro-biota. These dietary ingredients may act in an additive or a synergistic manner, exerting their effects on gut micro-biota. The prebiotic-like flavonol-rich foods have been demonstrated to modify the composition of gut micro-biota. Six week consumption of a wild blueberry drink that was high in polyphenols was shown to increase the proportion of Bifidobacterium spp. compared to the placebo group; however, a high inter individual variability in response to the dietary treatment was also observed.Similarly, the daily consumption of a high-cocoa flavonol drink for 4 weeks significantly enhanced the growth of fecal bifidobacterial and lactobacilli populations, but decreased the Clostridia histolyticum counts relative to those consuming a low-cocoa flavonol drink .

Furthermore, unabsorbed dietary phenolics and their metabolites selectively inhibit pathogen growth and stimulate the growth of commensal bacteria. For example, grape pomace phenolic extract increases Lactobacillus acidophilus CECT 903 growth in liquid culture media.In addition, upon bacterial incubation, tea phenolics were shown to suppress the growth of potential pathogens such as Clostridium spp. and Gram-negative Bacteroides spp., whereas commensal anaerobes such as Bifidobacterium spp. and Lactobacillus spp. were less affected.Similarly, the flavanol monomer -catechin significantly increases the growth of the Clostridium coccoides−Eubacterium rectale group, Bifidobacterium spp., and E. coli and significantly inhibits the growth of C. histolyticum group in vitro.To date, there is a wide range of phenolic compounds that have been demonstrated to have antimicrobial properties, and many have been previously reviewed.Although many of the studies highlighting the beneficial role of plant polyphenols through regulation by gut micro-biota appear promising, there are limitations in the results that can be drawn regarding the ability of flavonoids to influence the growth of selected intestinal bacterial groups using a batch-culture model. More comprehensive human intervention studies will be essential in the future to provide insight into the potential influence of dietary polyphenols and their aromatic bacterial metabolites on intestinal microbial communities and their activities.Probiotics are defined as viable microorganisms that, when consumed in sufficient amounts, confer a health benefit on the host.To date, most of the commonly used probiotics are limited to strains of certain Lactobacillus and Bifidobacterium species .

Survival during passage through the GI tract is generally considered as the essential feature for probiotics to preserve their active functions in the colon. Indeed, the probiotic strains must overcome biological barriers, including resisting gastric and bile acid secretion and tolerating intestinal lysozyme and toxic metabolites produced during digestion . Various studies found that at least a fraction of probiotic bacteria can be detected in stool for between 1 and 3 weeks after consumption . Probiotic Lactobacillus strains were also found to adapt for survival in the gut and possess gut-inducible genes that are responsive at different sites in the intestine.Interestingly, provision of the probiotic Lactobacillus plantarum to mice fed a Westernstyle diet and to humans resulted in similar gene expression profiles of this strain.As probiotics are delivered via various food vehicles, the complex food matrix should also be viewed as an important factor that may alter the probiotic activity in the gut. To date, only a few animal and clinical studies have addressed the functional roles of food on probiotic-conferred health benefits.The mechanisms of probiotic effects on health are only partially understood but likely function either directly through interactions with host intestinal epithelial and immune cells or indirectly by modulating the indigenous intestinal micro-biota. In regard to the latter, several studies have concluded that probiotic consumption does not result in global modifications of the intestinal micro-biota in healthy individuals.However, probiotics might confer modest but significant changes to the functional activities of local intestinal bacterial populations. When examined at the meta-transcriptional level, intake of a probiotic fermented milk was associated with the upregulation of microbial genes corresponding to plant polysaccharide metabolism.Similarly, administration of probiotics was shown to induce crosstalk between the probiotics from the diet and the individual bacterial species in the gut and might induce competition for limited substrates that results in fluctuations tof the metabolic profile of the host.The gut micro-biome of healthy adults is highly resilient , where the stable native micro-biota prohibits the succession of microbes from the diet.In addition, the effect of probiotics on the gut micro-biome appears to differ depending on host phenotypes such as age, health status, and chronic conditions. For example, the infant gut micro-biome is highly diverse and dynamically changes during development and therefore may be easily influenced by the consumption of probiotics . In individuals with irritable bowel syndrome , probiotic consumption resulted in an increase in the numbers of Bacteroidetes in the intestine.Moreover,hydroponic nft intake of two Lactobacillus strains by diet-induced obese mice altered microbial composition and decreased expression of inflammatory genes in the adipose tissue while increasing levels of fatty acid oxidation in the liver.Further studies are needed to investigate the effects of assorted probiotic supplements on the gut micro-biome with respect to various host life stages and phenotypes.

The premise behind substituting sugar with artificial sweeteners is to maintain the palatability of food at the same time as lowering energy intake. However, a sufficiently high ingestion of non/low-digestible sugar substitutes stimulates the growth of gut micro-biota and can induce transitory diarrhea in humans.In particular, the great proportion of non/lowdigestible sugar substitutes that reach the distal intestine are subject to fermentation by the colonic micro-biota, offering approximately 2 kcal/g of energy.Although discovering and characterizing these compounds within foods is relatively new, it is of interest to note that many of these food ingredients are common in our daily diet. For example, the disaccharide alcohol maltitol is considered a common replacement for sucrose. Urinary and fecal excretions of sorbitol and maltitol after 24 h in conventional rats were shown to be minimal compared with germ-free rats.Likewise, maltitol consumption significantly increased production of SCFAs in addition to nine tested fecal microbes after a 6 week trial, including bifidobacteria, Bacteroides, Clostridium, lactobacilli, eubacteria, Atopobium, Fusobacterium prausnizii, Ruminococcus flavefaciens, and R. bromii. A 12 week administration of Splenda, composed of 1.1% of the artificial sweetener sucralose, increased fecal pH and reduced the amount of fecal bifidobacteria, lactobacilli, Bacteroides, clostridia, and total aerobic bacteria in a rat, whereas isomalt, a widely used low-energy sweetener, was considered to be bifidogenic in a human study.Overall, artificial sweetener fermentation by gut micro-biota remains either unexplored or poorly documented, some of which are highlighted in a review by Payne et al..Azo compounds are widely used as coloring agents in foods, beverages, and food packaging.In addition, azo polymer coatings have been specifically designed for colon-selective drug delivery due to the presence of pH-sensitive monomers and azo cross-linking agents in the hydrogel structure.Indeed, azo dyes can be metabolized under anaerobic conditions by intestinal microbial processes and, as a result, produce the reductive cleavage products aromatic amines . The majority of the toxic effects of azo dyes are exerted through aromatic amines produced by their colonic degradation.Raffi et al. reported that isolated intestinal bacteria in an anaerobic culture system were able to decolorize the dyes in the supernatant, suggesting that some of the azoreductase activities are extracellularly released.Xu et al. demonstrated a variable degree of efficiency in the reduction of Sudan azo dyes and Para Red by 35 prevalent human intestinal microbes in vitro.In contrast, Sudan azo dyes and their metabolites selectively inhibit the growth of some human intestinal microorganisms,which may suggest a potential impact on gut micro-biome after long-term exposure. In summary, although there are tantalizing glimpses into the effect of azo dyes on microbes in vitro, more data from animal and human studies are keenly awaited.In the colon, sulfur is present in either inorganic form or organic form .The human GI tract poorly absorbs sulfate, and there is little sulfatase activity in the mucosa of the GI tract; therefore, free sulfate in the colon is likely to be of dietary origin.Dietary sulfate drives the activity of sulfate-reducing bacteria that couple oxidative phosphorylation with reduction of sulfate to produce sulfide.The total inorganic sulfur intake is much higher in the Western diet in comparison to a typical African rural diet. Highly processed foods that are high in sulfate include bread, soy flour, dried fruits, and brassicas, as well as sausage, beers, ciders, and wines.Sulfur-containing amino acids such as cysteine can be found in dietary protein and are a source of sulfur for colonic sulfate-reducing bacteria Desulfovibrio desulfuricans.Native Americans who consume a diet high in resistant starch and low in animal products harbor significantly distinct sulfate-reducing bacterial populations and more diverse and different methanogenic archaea than Americans consuming a typical Western diet.Substrate competition for hydrogen among methanogenic archaea, sulfate-reducing bacteria, acetogenic bacteria, and otherspecies likely occurs in the colon.Because hydrogen is an essential component for the survival of colonic methanogens, removal of the substrate terminates methanogenesis. Given an adequate supply of sulfate, sulfate-reducing bacteria that are more abundant in the right colon outcompete methanogenic archaea for H2 due to their higher substrate affinity to produce hydrogen sulfide , an end-product of dissimilatory sulfate reduction.As a result, the mucosal micro-biome may be shaped in part through the availability of toxic sulfide compounds and the differential susceptibility of mucosalistic microbes to the toxins.Furthermore, the activity of methanogenic bacteria can also be disrupted by bile acids. In brief, methane production was thought to occur only when sulfate-reducing bacteria were not active.If sulfate is limited and hydrogen is in relative excess, methanogenic bacteria or perhaps acetogenic bacteria will become essential.Therefore, the levels of sulfate present in the colon are critical for determining which bacterial group gains a better survival advantage.Alcohol. Many people consume alcoholic beverages; however, few studies exist on the effect of alcohol consumption on the gut micro-biome of healthy individuals. For individuals who consume alcohol to excess, abnormal gut micro-biota and bacterial overgrowth can potentially initiate or worsen alcohol induced impaired gut barrier function and contribute to endotoxemia in patients with alcoholic fatty liver disease.

An in vitro study, using a colorimetric assay, demonstrated a Brazilian date fruit could inhibit angiotensin-converting enzyme activity, a potentially important target mediating blood pressure both in the pulmonary circulation and endothelial cells. Results from in vitro oxidant defense assays have been further supported by ex vivo and in vivo animal studies. These studies demonstrated positive effects of different date cultivars against a variety of toxicants that produce free radicals, including carbon tetrachloride, isoproterenol , cadmium, and from the oxidant-generating streptozotocin-induced diabetic rat model. The protective effects of dates against oxidative stress were attributed to improved activities of oxidant defense enzymes such as catalase , superoxide dismutase , glutathione peroxidase, glutathione reductase, and glutathione S-transferase, along with a significant reduction in malondialdehyde. Moreover, dates were demonstrated to diminish oxidative damage, inflammation and apoptosis in cardiac tissue of ISO-treated rats. Oral administration of lyophilized Ajwa date extract down regulated the expression of pro-inflammatory cytokines and apoptotic markers in injured Wistar rat heart tissue, further supporting the anti-inflammatory and anti-apoptotic potential of dates against ISO-induced myocardial damage. A more recent study investigated the possible cardioprotective effects of a nanopreparation mix of Ajwa date fruits and seeds on doxorubicin -associated cardiotoxicity in Wistar rats by studying hemodynamic, electrocardiological, and biochemical change. The results obtained suggest that rats pre-fed 1.4 g/kg of the preparation one hour before DOX infusion were protected from a significant elevation in left ventricular pressure observed in the control group.

In addition, pretreatment with the preparation prevented DOX-associated ischemia and increased the antioxidant capacity of reduced glutathione in cardiac tissue,blueberry package compared with the untreated group. Quantification of Ajwa pits and a mixture of Ajwa fruit/pits reported that the pits contained 47.4 g/kg of total flavonoids, while the mix of pits with fruits contained 23.8 g/kg of total flavonoids, including epicatechin-phloroglucinol and epicatechin.Extracts of high concentrations of phenolic and flavonoid compounds in four different varieties of dates exerted favorable effects in protecting and repairing tissue injury following myocardial infarction induced either by ISO or temporary ligation of the left anterior descending coronary artery in a rodent model via oxidant defense activities and mobilization of circulating progenitor cells from bone marrow and peripheral circulation. In this regard, oral pretreatment with date extracts for a period of 28 days prior to ISO injection significantly improved the state of MI compared to the control group. Elevated levels of glutathione, SOD, and CAT, and reduced levels of thiobarbituric acid reactive substances in heart tissue of rats were noted. A dose-dependent effect was evident in which 400 mg/kg had significantly greater effects compared to 200 mg/kg in most cases. Interestingly, following MI induction, compared to controls, date extracts significantly increased circulating levels of CD34 and CD133 positive progenitor cells that are involved in tissue repair. The quantity of total phenolics in the dry plant material varied from 21.53 ± 0.90 to 26.82 ± 0.92 mg GAEs/g and from 2.90 ± 0.13 to 4.92 ± 0.21 mg quercetin/g.A limited number of studies in animals has suggested beneficial effects of dates on plasma lipids. Golden Syrian hamsters fed a high cholesterol diet supplemented with 50% Khalas date pulp for 13 weeks showed significant decreases in plasma cholesterol, triglycerides and LDL-C compared to those consuming the high cholesterol diet alone.

Interestingly, the addition of dates to the high cholesterol diet significantly increased liver triglyceride levels compared to the high cholesterol diet alone. Although the mechanisms underlying this liver lipid-loading effect are not clear, fructose, which is present in dates, has been shown to stimulate hepatic triglyceride synthesis when consumed with a hypercaloric diet. Unfortunately, insufficient information was provided regarding the composition of the chow and high cholesterol diets , or how liver cholesterol and triglyceride concentrations were measured, limiting the interpretation of the reported effects. In another study, hyperlipidemia-induced albino male rats fed either 300 or 600 mg/kg body weight of Aseel date fruit suspension for eight weeks showed significant reductions in triglycerides and very low-density lipoprotein compared to the control group. Curiously, serum levels of cholesterol, LDL-C, HDL- cholesterol and the LDL-HDL ratio were also significantly decreased at the intake of 300 mg/kg but not at 600 mg/kg. Results from the animals receiving date intake at 300 mg/kg approached the same response as the positive control group receiving atorvastatin . While intriguing, these data must be interpreted with caution, since rodents have a different absorption, distribution, metabolism and excretion profile for lipids compared to humans. A search of English- and Arabic-language databases revealed two human studies on date intake. One is a pilot trial with ten healthy non-smokers consuming 100 g/day of either Medjool or Hallawi dates for four weeks in a crossover design with a four-week washout between groups. A significant 15% reduction in serum triglycerides with Hallawi dates was noted compared to baseline values. However, the baseline triglyceride levels for those who consumed Hallawi dates were considerably higher than that of the Medjool group, although the actual values are difficult to assess since they are only presented by a bar graph.

The study also noted that the intake of Hallawi, but not Medjool dates, significantly reduced markers of oxidative stress, as measured by the TBARS assay, the 2,20 -Azobis hydrochloride-induced serum lipid peroxidation method, and increased serum paraoxonase 1 aryl esterase activity, an enzyme necessary to protect serum lipoproteins from oxidation. The beneficial reductions in oxidative stress after consuming Hallawi, but not Medjool dates, could be related to the total phenolic concentration, which was significantly greater in the Hallawi dates by 20% to 31%. Although the major proportion of the soluble phenolics in both date varieties consisted of phenolic acids, only Hallawi dates contained a significant portion of catechins, which have been reported to exert potent oxidant defense actions. Differences in absorption, metabolism, and bio-activity of the different phenolic compounds in the two date varieties may also help explain the disparate outcomes. Unfortunately, the inclusion and exclusion criteria for participants was poorly defined, and no data were provided regarding whether a return to original baseline values were noted after the washout and crossover period. Due to the high fiber content of dates, which can alter gut micro-biome profiles, the influence of a carry-over effect from the intake of one date variety to the next must be considered. Future study designs would ideally include a separate, no-intervention control group as well as a parallel arm design, or ensure that the primary lipid outcome measures along with the gut micro-biome milieu at the start of the second intervention period returned to original baseline values. A second human study assessed the effects of the daily intake of three Khudary dates for 16 weeks among 100 Bahraini adults with diabetes and control group consuming no dates [n = 50]. This randomized, controlled, parallel-arm study trial reported a significant improvement in the date group in plasma cholesterol levels of approximately five percent from baseline, along with a trend for reduced LDL-C. However, the results did not reach statistical significance when thetreatment group was compared to the control group, suggesting that some of the reduction in cholesterol may have been due simply to participation in the project, independent of date intake. Other limitations include lack of details regarding the actual weight of dates provided,blueberry packaging and the overall macronutrient distribution and quantity of the diets. Taken together, the reports above suggest that dates can improve markers of cardiovascular health, particularly plasma lipid levels, indices of oxidative stress and inflammation, and circulating progenitor cells. These results are primarily from in vitro and animal models, which may be useful as pre-clinical models. Unfortunately, differences in study design, the amount and composition of dates or extracts tested, and lack of details about the control groups fail to provide specificity and limit the ability to draw conclusions about mechanisms and applications to humans. The selection of a control item is important when investigating cardiovascular effects of date products or extracts, because controls can have significant amounts of bio-active compounds that can potentially affect cardiovascular function. Polyphenols are among the most studied categories of dietary phytochemicals in relation to vascular health, and the subgroups of flavanols, anthocyanins and proanthocyanins have been of particular interest. Intake of flavanol- and PAC-rich foods and food extracts from strawberries, blueberries, and cocoa have demonstrated improvements in vascular markers that are associated with markers indicative of improved cardiovascular health. Several molecular mechanisms contribute to the physiological effects of flavanols, including enhancement of vasodilation through the induction of NO , free radical quenching , inhibitory effects on select prooxidants , and reduction in ET-1 activity.

Effects of dietary flavanols on markers of cardiovascular health have been discussed in detail elsewhere. Date cultivar and degree of ripeness are major determinants regarding their polyphenolic composition, as discussed above. Phenolic-acid and flavonol fractions isolated from Amari and Hallawi dates at the tamer stage were examined in vitro for antioxidant and antiatherogenic properties. The two fractions exhibited variable capacities to reduce ferric ions , scavenge radicals and inhibit LDL-C oxidation via TBARS and lipid peroxide assays, with the flavonol fractions showing the strongest effects. Only the flavonol fractions stimulated cholesterol removal from macrophages. The yields of isolated fractions in terms of µmol GAE per g fruit were considerably larger for Amari dates by approximately 10- and 3.5-fold for phenolic acids and flavonols , respectively. The two isolated fractions contained ferulic acid as a major component and comparably small amounts of coumaric acid, but differed considerably in the composition of their complementary set of phenolic acids. Amari dates contained primarily caffeic acid derivatives, whereas the Hallawi variety contained mostly a salicylic acid derivative. Seven prominent peaks of flavonols were evident. Based on an authentic standard library, all seven flavonols were tentatively classified as kaempferol derivatives. The two isolated fractions of date flavonols differed considerably in composition. In addition to one prominent flavonol peak shared by the two fractions, Amari consisted of significant amounts of five other flavonols, whereas Hallawi contained a single unique flavonol as the major component. The results demonstrated strong structure-activity relationships for date polyphenols and identified date flavonols as potential antiatherogenic bio-actives. Polyphenols derived from date syrup at concentrations of 60 and 600 µg/mL, predominantly from cinnamic acid and catechin derivatives, were found to significantly attenuate IL-6, IL-8 and vascular endothelial growth factor in human vascular endothelial cells. These observations corresponded to asignificant reduction of both cyclooxygenase-2 and VEGF induced by TNF-α at both the protein and gene expression levels in the assessment of inflammatory-associated angiogenesis in HECVs. Many polyphenols found in dates have been studied as isolated compounds in in vitro and ex vivo systems with respect to their effects on markers of vascular function. Protocatechuic acid, a metabolite of the anthocyanin cyanidin-3-glucoside , and its phase II metabolites were effective in modulating the production of the key inflammatory mediators IL-6 and vascular cell adhesion molecule-1 at dietary-relevant concentrations as low as 100 nmol L−1 , with maximum reduction observed for the sulfate conjugates in human umbilical vein endothelial cells stimulated with either oxidized LDL or a cluster of differentiation CD40L . In the same study, C3G and its metabolites reduced IL-6 production in CD40L-stimulated cells, whereas both C3G and its metabolite, ferulic acid, reduced VCAM-1 production. Anthocyanins and ferulic acid have also been found to significantly reduce monocyte adhesion to HUVECs under physiologically relevant conditions, an important step in reducing atherosclerosis development. In human intestinal cells in vitro, the addition of a freeze-dried date extract from California-grown dates with a total proanthocyanidin content of 13% was demonstrated to act as a potent co-agonist ligand for the farnesoid x receptor , a nuclear structure important for maintaining triglyceride and cholesterol homeostasis. This study provides a potential mechanism by which dates may exert a hypotriglyceridemic effect, as observed in the human study noted above. The tea catechin, epigallocatechin- 3-gallate , has also been shown to modulate FXR in a tissue- and gene-specific manner. Further studies are warranted with dates and their extracts using both wild-type and FXR knockout mouse models. While the above in vitro work is promising, data from dietary interventions that specifically examine the association between circulating date polyphenols or phenolic metabolites with physiological effects have yet to be reported.

Starch, or the proportion of the amylose fraction of starch, is used as a common ripening biomarker for apple, banana, and pear. This marker relies on the ability of amylose to physically interact with iodide to form a triiodide blue-black complex. Starch can also influence the quality of fruit juice. Although starch is degraded to sugars when fruit ripens, this conversion is not complete. Ripe fruit processed for juice therefore contains starch, which is treated with amylases for clarifcation. Further, the amylose content of the remanant starch in some fruit processed for juice, may alter juice viscosity.Prepackaged leafy greens are convenient and healthy, and are popular options for salads in western countries. Metabolism in this horticultural product can be considered over distinct phases in its lifecycle: pre- and post harvest. In developing spinach, the photosynthetic organ, i.e., the leaf, fixes carbon, and partitions a large portion ~20% to starch biosynthesis during the light period under lab conditions . During the night, the leaf starch is degraded into sugar, to maintain plant metabolism, resulting in an empty polysaccharide reserve before the next light period. In Arabidopsis, the expression of SBEs and the changes of amylopectin and amylose show a similar trend, but there is variation in when SBE transcripts peak. Although there is no information on SBE transcriptional levels in spinach during the diel,nursery grow bag there may be some similarities with Arabidopsis because the pattern of leaf starch accumulation is comparable in spinach and Arabidopsis. The preharvest starch reserve may alter the postharvest quality of leafy greens.

Harvested green produce are stored in optimized packaging under limited light exposure conditions which restricts new energy and carbon input from photosynthesis. However, respiratory activity, which is the carbon skeleton generation process for cellular metabolites, although reduced, does not stop. In detached leaves, the starch can be broken down to glucose, and sugars become the main source of fuel for cellular metabolism and ATP generation in the early stage of respiration . In the late stage of the respiratory process, the depleted sugars will be replaced by proteins, lipids, and membranes, triggering leaf senescence and cell death. Tis results in undesirable produce quality and ultimately, in produce loss. Preharvest and post harvest starch content may determine post harvest energy reserves and influence the time span that buffers the onset of senescence, thus influencing shelf-life of harvested green leaves. Correlations between leaf starch content and post harvest longevity have been found. For example, lettuce and red chard harvested at the end of the day, when leaf starch content was highest, had a longer extended shelf-life than organs harvested at other times of day. This may not be true of all varieties e.g., salad roquette. Starch also correlated with improved shelf-life quality after light exposure to detached leaves in vegetables such as Chinese kale and lettuce. The accessibility of sugars from the degraded starch may relate to leafy-green quality, and the upregulation of SBEs would convert amylose to the more catabolically available amylopectin, providing a more readily available source of sugar.The amylose-to-amylopectin ratio in Arabidopsis influences flowering time and reproductive growth, key markers of development, and fitness. Whether starch molecular structure and composition influences the preharvest growth of leafy greens in a similar way, remains unknown, but it seems likely.

Potato, sweet potato, and cassava are generally considered as high glycemic index foods because the starch in their storage organs is easily digested to sugars when consumed, leading to a rapid increase in blood sugar level. It is established that high GI food exacerbate metabolic disorders such as diabetes and obesity. In contrast, multidisciplinary experimental research shows that digestion-resistant starch could increase the healthful microbial communities of the gastrointestinal tract, reducing the occurrence of constipation, and lowering the risk of colon cancer. Altering potato starch composition is a viable way to increase ‘dietary fiber’ content and to enhance colonic health. This can be achieved by either physical, chemical, or enzymatic modifications of purified starch, e.g., etherification, esterification, or by fine-tuning the activity of starch biosynthetic enzymes. Reduction or knockout of SBEs in a range of species have reliably led to an increase in the resistant starch content in various species including horticultural crops e.g., potato, sweet potato, and cassava. Interestingly, SBE2 is not the dominant isoform expressed in storage tubers and roots, but it exerts a major function in amylopectin synthesis. Very high levels of RS can be achieved by the combined suppression of SBE1 and SBE2, but with a yield penalty. The transcriptional profiles and functions of SBE3 are unclear in the developing tubers . In addition, potato tubers suffer from a post harvest disorder: cold-induced sweetening . Potato tubers are stored at low temperatures to extend shelf life and to meet year-round demand. However, sugars accumulate from starch breakdown, a process referred to as CIS. Although a problem for the potato industry, CIS could be a mechanism to allow tubers to cope with chilling stress. CIS negatively affects the quality of fried or baked potato products: reducing sugars react with free amino acids at high temperature cooking through the Maillard reaction, to form carcinogenic acrylamide.

Changes in the enzymes involved in starch biosynthesis and degradation are involved in CIS. SBEs are actively expressed in CIS susceptible tubers, and in StVInvsilenced, CIS-resistant tubers, SBEs transcriptional level were suppressed. Naturally occurring high RS potato varieties, also, have less susceptibility to CIS. Therefore, evidence points to a positive association of SBE activity with CIS severity in some potato genotypes.Starch is a major component of the dry mass of fruits at commercial harvesting time. Starch is transiently synthesized and stored in unripe fruits with a peak just before ripening. Starch appears to be a critical feature of climacteric fruit metabolism, known for their bursts of respiratory activity and ethylene production upon ripening. Climacteric fruits contain more starch, and, more active starch biosynthesis than non-climacteric fruit after anthesis. In tomato, the functional genomics model for freshy climacteric fruit, starch fulfilled 40% of the carbon needed for respiratory processes based on a constraint-based flux model. Experimental evidence from post harvest metabolism also supports the model: tomato fruits stored post harvest under low or chilling temperatures undergo bursts of stress-related carbon dioxide and ethylene production when allowed to recover at room temperature, with an accompanying and corresponding decrease in starch reserves. A similar inverse relationship between starch content and respiratory activity was observed in ripening banana, ginger rhizomes sunberry, apple and durian. Te relationship between tissue starch content and respiration may not be perfectly linear in all species, e.g., in stored ginger, starch showed a biphasic accumulation pattern as respiration progressed, a trend not seen in other tissues examined . Furthermore, the relationship between these variables may also differ among genotypes within a species. Apart from climacteric characteristics, after the onset of ripening, starch content plummets sharply accompanied by starch decomposition into soluble sugars, and the total soluble sugar content continues to rise proportionally . This dynamic metabolic process had been reported for both climacteric and non-climacteric species including tomato, apple, banana, plantain, mango, kiwifruit, pear, and strawberry. Adequate storage of the starch-derived soluble sugars, is essential to produce an acceptably favored horticultural produce of appropriate sweetness. Accompanying the starch-sugar dynamics,plastic growing bag amylopectin-to-amylose ratio , also changes interactively . The difference in the AP/AM ratio in fruit development is expected to influence the structure of starch and its degradability. In the ripening tomato, the rate of decrease of amylose was greater than that for amylopectin . Thus, the AP/AM ratio increased dramatically during ripening, in concert with the increase in soluble sugar content and fruit color change from green to red. This phenomenon where the proportion of amylopectin increases relative to amylose, was also evident in ripening apple and banana. It is possible to speculate that of the available starch left during fruit ripening, the amylose, or long chained amylopectin was converted into amylopectin whose branch-like structure has a much higher susceptibility to enzyme attack, allowing the rapid process of starch degradation into soluble sugars and supply for respiration. However, this mechanism may not be universal for all fruit. For example, the changes in AP/ AM ratio in kiwifruit are similar to those in developing potato tubers, where the ratio of AP/AM almost remains constant during tuber development. In ripening tomato fruit with sharp increases in AP/AM, up-regulation of SBEs transcriptional expression is expected. Among SBEs, the class 2 SBE has the major effect on altering starch compositions .

Elevated expression of SBE2 transcripts does parallel the changes in the AP/AM in ripening tomato, apples, and banana. We propose that ultimately, this change in glucan structure indirectly contributes to favor, quality, and commodity value.Starch, in general, plays an essential role in balancing the plant’s carbon budget as a reserve of glucose that is tightly related to sucrose metabolism and sugar signaling pathways. Starch is considered as an integrative mediator throughout the plant life cycle, regulating plant vegetative growth, reproductive growth, maturation and senescence, and response to abiotic stresses. This comprehensive regulation is achieved by changes in the synthesis and degradation of starch to balance glucose levels, after developmental and environmental triggers in different organs. Transitory starch and its biosynthesis have been well studied in the model plant Arabidopsis, but little research has been conducted on post harvest leafy greens. Quality metrics such as shelf-life, favor, color, firmness, and texture are of consumers’ choice, and they are related to the limited pools of storage compounds in detached leaves, which cells rely on to maintain basic cellular activities. A hypothesized function for the starch in packaged leaves could be presented as such: starch may act as a bufer against sugar starvation, and protect against cellular autophagy, by serving as an alternative energy source. If the biosynthesis and degradation of starch could be adjusted in a controlled way, then the modulated release of sugars may influence the post harvest shelf-life in detached leafy greens . A continuous, paced supply of sugars may preserve vacuolar nutrients and water content, leaf cellular structure and integrity, and, thus extend the ‘best by’ post harvest date of the produce. Although the eco-physiological role of amylose is poorly understood in Arabidopsis, the AP/AM ratio may set a threshold for the optimum usage of starch. SBE action in leafy crops may differ from those in Arabidopsis given the dissimilar numbers of their isoforms and domain features . Modifying the quantity and quality of the starch in leafy greens such as spinach, lettuce, and watercress, by targeting starch bio-synthetic enzymes, may provide evidence to its post harvest function in terms of produce longevity. Resistant starch is a popular nutritional additive to produce food with enhanced quality attributes, i.e., higher fiber content, and starchy horticultural commodities are similarly attractive. Te yield penalty of high amylose crops may be alleviated by picking an ideal AP/AM ratio through a coordinate change in the relative balance of starch bio-synthetic enzymes. In the case of potato, it is plausible that down regulation of SBEs not only produces healthy fiber-starch, but also lessens the CIS severity and acrylamide problem . However, the sugars derived from starch during CIS may be an adaptive mechanism to enhance plant chilling tolerance. Rapid sugar accumulation upon cold stress have been reported in fruit . The sugars freed from starch may promote metabolic activity and serve as an osmoprotectant, thus alleviating chilling injury. Te major functional SBEs were found to be upregulated in cold-stressed banana fruit, potato tuber, and Arabidopsis leaf, which may facilitate the ‘sugaring’ process. Modulating SBE activities may alter the rate of sugar released from the highly digestible starch polymers, thus changing the fruit/tuber cold responses. In fruiting species, the importance of ‘transitory-storage starch’ may be underestimated due to the lack of enough direct knowledge of its function, gained from experimental data. Tomato serves as a functional genomics model for feshy fruit, as it is easily transformed and genetically manipulated. The putative function of ‘transitory-storage starch’ in fruit ripening, respiration, and sweetness enhancement may be revealed by engineering AP/AM ratio through over expression or suppression of SBEs. We hypothesize that high amylose, resistant starch tomato fruit may have reduced available starch, sugars, and changes in fruit ripening and other processes that are dependent on starch as a carbon supply and source of energy post harvest. Tomato SBEs may not reffect the functionality of all fruit SBEs, but it would produce fundamental knowledge and expand our understanding of species-, organ- and developmental-specifc regulations of the core starch bio*synthetic enzymes.

Our study showcases the value of field-based research in understanding diapause as a potentially vulnerable or robust stage in an insect’s life history.Lab-based studies that monitor bumblebee vital rates under controlled conditions will remain foundational in the effort to identify the environmental factors which impact diapause survival. However, lab-based studies cannot capture the real-world environmental pressures faced by diapausing insects, and lab-reared colonies cannot replicate the adaptive capacity of wild colonies to cope with these pressures. In demonstrating the utility of B. impatiens as a model system for exploring insect overwintering ecology, we are hopeful that our research will pave the way for future field-based studies targeting this life cycle stage.Our cultural context shapes our interactions with the world and dictates the way we visualize, process, and utilize information. For example, holistic and analytic modes of thought differs across cultures , with people from East Asian, collectivistic cultures typically exhibiting holistic thought and people from Western, individualistic cultures exhibiting analytic thought . Holistic thought associated with an East Asian perspective is primarily dictated by context and relational cues. In this style of thinking, people are more likely to attend to background information and relationships, viewing the system as a whole rather than a sum of its parts . For example, in the classic narrative task, participants view an image of a fish bowl and describe what they see in as much detail as possible for one minute . Holistic thinkers, in this task, are more likely to name background or environmental objects like “under the water.” Analytic thought, in comparison,black flower bucket is characterized by attention to foreground information and discriminating/categorical qualities .

In the narrative task, analytic thinkers are therefore more likely to name foreground objects like “fish” and describe their properties and movements. Previous research has predominantly focused on collectivist and individualist cultural differences in different thinking styles, and factors that could explain the culturally bounded phenomenon . For instance, researchers have examined the role that environment plays in perception, exposing participants to images of cityscapes . These researchers found that images of Japanese cities were more ambiguous and contained more elements than images in American scenes, presumably priming participants to attend more carefully to context. Indeed, when primed with images of a Japanese city, both American and Japanese participants were more likely to think holistically. In addition to environmental differences, language might also play a role. For instance, in one study, researchers examined whether thinking styles influence attention and awareness. To test this, participants were shown the classic fish bowl vignette, and researchers used an eye-tracking device to measure eye fixation patterns. East Asian and Western participants showed non-significant differences in looking patterns . However, the emergence of holistic and analytic thoughts occurred when participants were asked to describe the scene, where East Asian participants were more likely to name background and environmental information than Westerners did. Since differences were only observed when participants were asked to describe the image, researchers suggested language as a mechanism for divergent thinking styles. Language differences could explain sub-cultural differences as well. Two studies found differences even between two collectivist cultures. Korean participants thought more holistically than Chinese participants did when presented with the narrative task. Researchers proposed that these varying degrees of holistic thinking might be related to the syntactic differences in the participants’ native languages, noting that English sentences were mostly head-initial and Korean were mostly head-final.

This “head-initial/head final” judgment refers to verb/noun positioning, with head initial language leading with the noun followed by the verb, and head-final language leading with the verb followed by the noun. This hypothesis suggests a primacy effect, with speakers being drawn to whatever aspect of the sentence is placed first. English, therefore, as a head-initial language, would mention the subject first in a sentence, priming its speakers to attend to focal objects, whereas Korean, as a head-final language, would mention the subject last in a sentence, priming its speakers to attend to context and the relationship between objects. Since Chinese alternates between head-initial and head-final languages, Chinese speakers showed a lesser degree of holistic thinking than Korean speakers. Results from other studies support the idea that language properties could associate with the measured thinking styles across individuals, situations, cultures, and even subgroups within the same culture . So far, the literature has suggested thinking styles as a static characteristic of members of a culture, and unique features of language could foster a certain type of thinking style over a long-term exposure. Little is known how language could potentially influence thinking styles as a contextual factor from moment to moment. For example, studies have used tasks with categorical or relational associations of concepts to measure holistic and analytic thinking styles. One influential study constructed triads with both a categorical and relational choice. Participants were asked to choose the two words that were most closely related . If the participant grouped policeman and postman, this would be categorical, indicating an analytic choice, whereas policeman or postman grouped with uniform would be relational, indicating a holistic choice . However, ample studies have shown that the strength of categorical and relational associations can vary across pictorial and verbal formats . In general, pictorial stimuli yield a stronger categorical association based on semantic feature overlaps, whereas verbal stimuli yield stronger relational associations based on word associations. Consequently, the verbal triad could “benefit” relational associations more whereas the pictorial triad could “benefit” categorical associations. Therefore, it is possible that the measured thinking style could be dependent on the format of the triad task. Furthermore, categorical associations could be manifested in the verbal labels of object names. For example, some semantic categories are emphasized in their names, such as “berry ” and “nut ”.

Having these verbal labels in object names could potentially emphasize the categorical associations of concepts, influencing the thinking styles in different ways. On the one hand, since the categorical association is related to analytic thinking in the triad task, highlighting the categorical information could lead to more analytic thinking . On the other hand, highlighting the categorical information via verbal labels of object names could potentially promote a focus on similarities of objects, a feature of the holistic thinking style . Anecdotal evidence is in alignment with this argument. Specifically, this feature is extremely pervasive in some Asian languages, such as Mandarin Chinese, but not so much in Western languages such as English. Chinese names of objects commonly include the category label alongside the noun, such as the verbal label “flower ” in “rose-flower ”, “daisy-flower ”, and “peony-flower ”. Potentially, having this language feature in Mandarin Chinese promotes a focus on overall similarities of objects, fostering holistic thought in Chinese speakers. The overarching goal of this study is to examine how language might impact thinking styles, by using verbal labels to create different contexts when measuring people’s thinking styles. Study 1 sought to examine whether thinking styles measured in a classic triad task could be shifted using verbal labels instead of pictures. Given the evidence that relational associations are stronger in verbal format,square black flower bucket we anticipated that people would show more holistic thinking when tested with verbal format in comparison to pictorial format. Study 2 aimed to examine whether participants could shift their thinking styles measured by the verbal triad task, after being verbally primed to focus on categorical associations of objects. Two potential outcomes were predicted according to two competing hypotheses . If the effect of verbal labels is direct , we would expect that the categorical labels in object names would make people focus on the categorical associations of words, and choose categorical associations more . However, if the effect of verbal labels is indirect, we would anticipate that exposure to these category labels in object names could make people focus on the overall similarities of objects, and they would show a shift to holistic thinking and choose more relational associations. In addition, we explored potential gender differences in measured thinking styles since studies have shown that females and males could differ in their analytic skills and intuitive-analytic thinking .The different thinking styles, namely, analytic vs. holistic, have been long associated with Western and East Asian cultures, respectively. Our findings from two studies have provided some preliminary evidence that thinking styles of native English speakers, especially when measured by verbal formats, are highly variable across individuals and context dependent. In Study 1, we showed large individual differences in thinking styles, and when measured with a verbal format, more individuals became holistic thinkers from analytic thinkers compared to when measured with a pictorial format. In Study 2, we used an experiment to demonstrate that exposure to verbal labels highlighting the categorical associations would lead participants, especially females, to prefer categorical candidates in the verbal triad task. These findings converge on the idea that thinking styles can be variable instead of static, and language can impact thinking styles of individuals from moment to moment.Results from Study 1 and Study 2 may seemingly be contradictory since Study 1 suggested that people became more holistic thinkers with the verbal triad task whereas in Study 2, participants were shifted to analytic thinkers after being primed with verbal labels.

However, they actually converge on the observation that language could impact thinking styles, but the exact direction depends on the feature of the verbal information and the contexts. The triad tasks in Study 1 assess relative strength of categorical and relational pairs of images or words, resembling word association and semantic priming . Previous meta-reviews have pointed out that both semantic and relational associations could lead to a priming effect, but the semantic association depends on feature overlaps of objects whereas the relational association is also influenced by word association in text . In addition, a previous study has shown that the semantic priming effect is stronger in the verbal than pictorial formats . Therefore, when the triad task was implemented in a verbal format, the relational association became “boosted” so people chose the relational candidates more compared to the pictorial format. In Study 2, we primed people to focus on the categorical associations by using names that highlighted the categorical information. Specifically, the verbal labels like “berry”, “nut”, and “ball” in the prime condition made the shopping task easier to the participants if they looked for the verbal similarity. This verbal similarity would then further promote a focus on the category similarity. Consequently, participants were potentiated for the categorical association in the triad task. Thus, the influence of the verbal labels was directly “translated” into the triad task, and people tended to continue focusing on the categorical associations and chose the categorical candidates more, resulting in a shift to analytic thinking. The different results of Study 1 and Study 2 could also partially, if not all, result from the task instructions. In Study 1, we instructed participants to find candidate word/picture that goes “together” with the target word, indicating an emphasis on relationships; however, in Study 2, the shopping task required participants to “sort/separate” items, promoting some analytical processes. Nevertheless, this possibility still points to the factor that thinking styles can be viable and influenced by language. Therefore, although the influence of verbal labels shifted people’s thinking styles in the opposite directions in Studies 1 and 2, they both manifested the potential impact of language on measured thinking styles. It is worth noting that our findings mostly showed the short-term effects of language on thinking styles, since the verbal labels only created task-specific contexts when measuring thinking styles. At most, our results suggest that language could influence thinking styles and explain cultural differences in thinking styles . In particular, we want to focus on the results from Study 2. One motivation for Study 2 came from the observation that categorical associations are prevalently emphasized in Mandarin Chinese, so this feature in Mandarin Chinese could potentially cultivate a focus on the overall similarity of object categories, fostering a holistic thinking style. In contrast to this hypothesis, we did not observe that people became more holistic thinkers after being primed with categorical information. Instead, we found that categorical information made females in our sample more likely to use analytic thinking. Hence, in a Western sample, categorical information might have a direct effect, because it makes people focus on the primed category and its distinctiveness from other categories more.

The leaching water was collected according to the precipitation. Gas fluxes at the soil surface were sampled by the closed chamber technique, and N2O content was analyzed using a Shimadzu GC-14B. Drainage water and soil NO3 – -N concentrations were determined by the Hydrazin-reduction method . The pH of soil and drainage water was measured using 1:2.5 soil to water ratio and a Horiba D52 pH meter. The EC was measured using a 1:5 soil to water ratio with a TOA CM-14P EC meter. Leaf chlorophyll content was monitored using a SPAD meter . The NH4 + concentration of soil immediately responded to ammonium sulfate fertilization, particularly in the SC treatment. The ammonium sulfate input resulted in a higher NO3 – concentration due to nitrification. Supposedly, the SC and PM treatments had reduced nitrifier activity, thus the soil NO3 – peaks appeared one month after the SO treatment. An earlier peak of NO3 – concentration in the month of fertilization in SO indicates higher activities of nitrification compared with other treatments. The highest NO3 – concentration occurred in the PM treatment in August 2008. The pH patterns of soil strongly correlated with the amount of precipitation and fertilizer applications. The low cation content of the rainwater may enhance possibilities of leaching base cations in soil out of its profile due to their replacement. The high rainfalls in June 2008, coupled with ammonium sulfate fertilization, apparently introduced an acidifying reaction in the soil solution. The soil disturbance soon after the plantation of blueberry bushes to the containers generated a more aerobic and suitable environment for mineralization,plastic flower bucket resulting in the release of NO3 – in the early stage of the experiment.

The SC treatment originally contained high N contents derived from sewage sludge, which resulted in higher NO3 – losses in May 2008. The nitrification induced nitrate loss occurred after the first ten days of August. The SC treatment had no leaching water released from the container in the second decade of August, probably because of the dense root mat having a very active water and nutrient uptake. The other possible reason is the fast root elongation in the phenological stage around the middle of August . The total NO3 – losses of PM, SC and SO treatments were 275, 475 and 453 mg, respectively. The gradually decreasing EC patterns indicated strong leaching of base cations from the soil profile. This cation impoverishment of the media gradually became moderate in the first decade of August when the fertilizer treatment resulted in a higher cation release from the soil cation exchange sites. This fertilizer induced higher EC stabilized around the first decade of September. The plant transplantation to the containers increased the soil pore space, thus providing more aerobic conditions for microbial activity, and resulting in higher NO3 – leaching. Higher N2O emissions were detected in SC before fertilization. These may have been derived from the initially high NO3 – content in the amended material. The emission factor in SC was 0.73%. After fertilization clear fertilizer-induced N2O emissions were detected in SO and PM treatments. The emissions factors of SO and PM were 1.2 and 1.43 %, respectively. The SO represents a typical heavy textured soil widely known as having physical characteristics to emit higher N2O, compared to the light textured ones . The SO treatment was under a stronger anaerobic condition for longer periods than in the PM and SC and resulted in higher N2O emissions.

Surprisingly, the light textured PM had also high fertilizer-induced N2O flux. In general, an increase in acidity through various mechanisms may inhibit the N2O flux. This may be related to the highest acidity of the media and the acidifying effects of fertilizer. As a recent study pointed out, the contributing microorganisms for N2O emissions may be more abundant than previously known . Especially, the tropical and boreal peat soils have a wide fungal diversity including denitrifiers. Peat moss as a soil amendment might be a potential source of fungal denitrifiers, which finally results in higher emission rates of N2O. In conclusion, the highest loss of NO3 – was obviously generated by the soil amendment application and plant transplantation. For as much as the blueberry is a perennial crop, this soil disturbance and heavy application of organic matter apply only in the establishment year. The subsequent years of plant management can be carried out without any deep tillage or organic amendment application that may release an excess amount of NO3. Taking into account the above mentioned characteristics of soil-plant-water-gas relations, the sawdust compost amended treatment has the least environmental impact in upland soil cultivation of blueberry. Over the past decade, the number of H-2A guest workers employed on California farms increased more than tenfold, so that almost 44,000 farm jobs were certified to be filled by H-2A workers in fiscal year 2022 . During FY2020, two-thirds of the H-2A jobs certified in California were in crop support services. Most crop support jobs were with farm labor contractors , but one sixth were hired directly by fruit producers. Almost 10% of the H-2A workers were hired directly by vegetable producers . Until the 2008–2009 recession, most H-2A workers were in southeastern states such as Florida.

However, the slowdown in migration of undocumented individuals after 2008–2009, combined with a stable demand for farm workers and the aging and settling of undocumented workers who arrived before 2008–2009, contributed to the rapid growth in the H-2A program in the three Pacific Coast states that employ half of U.S. farm workers, a third of whom work in California.This paper analyzes agricultural employment data from the California Employment Development Department and the Quarterly Census of Employment and Wages to understand changing patterns of farm worker employment in the 21st century. The data show that seasonality is declining in most regions and commodities, primarily because of higher employment during the winter months, which may reflect more winter pruning jobs and fewer summer harvesting jobs. Second, the data emphasize the increasing importance of non-farm crop support employers, mostly labor contractors, who bring workers to farms to perform specific tasks. More farms appear to be developing a year-round workforce that is hired directly and supplemented when needed with workers brought to farms by labor contractors, including H-2A guest workers . California requires all employers who pay $100 or more in wages to enroll in the state’s unemployment insurance system and pay taxes of 1.5% to 6.2% on the first $7,000 of each employee’s wages to cover the cost of unemployment benefits for laid-off workers . Employers also report their employment for the payroll period that includes the 12th of the month. Summing these monthly employment totals and dividing by 12 months generates average employment, also referred to as year-round equivalent jobs. The monthly employment measures allow us to determine the peak and trough employment months. Agricultural employment, as defined by the North American Industry Classification System , peaked at 470,000 in May 2020 and was 346,000 in March 2020, generating a peak-trough ratio of 1.4. More than 470,000 workers are employed on California farms sometime during the year. Workers who are employed only in payroll periods that do not include the 12th of the month, such as those who work only during the first, third, or fourth weeks of the month,flower buckets wholesale are excluded from average employment. In 2016, when California’s agricultural employment averaged 425,000, almost a million unique Social Security Numbers were reported by the state’s agricultural employers, suggesting 2.3 unique workers for each year-round equivalent job .California became the leading farm state in terms of sales in 1949, when Los Angeles County led the United States in farm sales . The state’s population doubled between 1950 and 1970, from 10 million to 20 million, and agricultural sales grew fastest in the San Joaquin Valley after water projects allowed more acres to be irrigated and suburbanization reduced the availability of farmland in coastal areas. Citrus and dairy farms in Southern California migrated north to SJV, while tree fruit farms moved from the urbanizing Bay Area to the San Joaquin and Sacramento valleys . Three SJV counties — Fresno, Kern and Tulare — accounted for 20% of California farm sales in 1949, a third in 2000, and almost half of the state’s farm sales in 2020 . Some crops that were already concentrated in the Sacramento and San Joaquin valleys expanded in acreage. For example, there were 90,000 bearing acres of almonds in 1950, almost 150,000 acres in 1970, 500,000 acres by 2000, and 1.3 million acres in 2022. Most of this additional almond acreage was in the San Joaquin Valley.

New orchards and dairies in the San Joaquin and Sacramento valleys were often larger and more efficient than the coastal farms they replaced, and their higher productivity was reflected in rising yields. Average yields of many fruits and vegetables doubled and tripled over the past three decades; bell peppers and cantaloupes are examples. Yield rose over 50% to 33 tons an acre for strawberries . The major change in California crop farming over the past half-century has been the rising share of high value fruits, nuts, vegetables, and melons, as well as horticultural specialties such as flowers and plants, in the state’s farm sales. In 1960, the value of fruit, vegetable, and horticultural commodities was two-thirds of the total value of California crops; since 2000, FVH commodities have accounted for over 90% of the value of California crops, reflecting growing consumer demand for fresh produce and nursery plants . Cotton was California’s most valuable crop in 1950; by 2000, cotton was the sixth most valuable crop, and by 2020 cotton was no longer among the state’s top 20 crops. The demand for FVH commodities rises with income, and rising farmland prices encouraged individuals and investors to buy farmland as a hedge against inflation in the 1970s, a decade in which the value of California farmland more than doubled . Higher interest rates in the 1980s led to a farm financial crisis that was more severe in mid-western states than in California, but encouraged some oil firms and conglomerates to sell their California farmland. The data in table 1 show that California’s farm sales almost tripled in three decades, and that fruit and nut sales almost quintupled. The value of the state’s vegetables and melons doubled, as did the value of greenhouse and nursery crops. The state’s farm sales were $17.8 billion in 1990, including $4.4 billion worth of fruits and nuts and $3.9 billion worth of vegetables. Farm sales were $27.2 billion in 2000, including $7.3 billion worth of fruits and nuts, $6.2 billion worth of vegetables, and $2.8 billion worth of greenhouse and nursery commodities. This rose to $37.5 billion in 2010, including $13.5 billion worth of fruits and nuts, $6.7 billion worth of vegetables, and $3.8 billion worth of greenhouse and nursery commodities. In 2020, farm sales were $49.1 billion, including $20.6 billion worth of fruits and nuts, $7.8 billion worth of vegetables, and $6.3 billion worth of greenhouse and nursery commodities. In real or inflation-adjusted terms, California farm sales rose by 40% over 30 years, and fruit and nut sales by 140%, while vegetable and nursery sales were little changed.Many FVH commodities are labor intensive, so expanding production increases the employment of farm workers. Rather than hiring workers directly, many farmers are turning to crop support service firms. These are non-farm businesses that bring workers to farms to accomplish specific tasks. For example, farmers may rely on labor contractors to bring crews of workers for a few weeks to prune, thin, or harvest their crops. Contractors may be the sole employers of the workers they bring to farms under some labor laws, such as unemployment insurance and workers’ compensation, and joint employers with farms under others, such as the Agricultural Labor Relations Act. Over the past three decades, California farmers hired 20% fewer workers directly, reducing average direct-hire employment in crops from 203,000 to 160,000. Meanwhile, crop-support employment rose by 60%, from an average of 132,000 in 1990 to 212,000 in 2020. Combined crop and crop-support employment accounts for over 90% of California’s agricultural employment. Within crop support employment,the farm labor contractor share of average crop-support employment rose from 60% to 67% . FVH commodities account for 90% of direct-hire crop employment, including 55% for fruits and nuts, 20% for vegetables and melons, and 15% for greenhouses and nurseries.

A modified version of this model was used to analyze the data from the four orchards with observations at both high and low wind speeds, to explore a possible interaction between the frequencies of flower visits in different sections of the tree, under different wind conditions. The model was the same as previously described with the orchard’s pollinator diversity category, the wind level, the location within the tree, and their interactions included as explanatory variables. To explore any differences in the spatial patterns of the four taxa under low and high wind speeds, only the data from the two high diversity orchards were selected. The same model as above was used with the wind level, the location within the tree and their interaction as the explanatory variables. For all models, stepwise deletion was carried out and an ANOVA comparison made between the model with and without the variable as previously described. The aforementioned analyses of flower visitation in high and low wind speeds were repeated only using the data from 2008. All analyses were carried out in R version 2.14.1 .Here we show how pollinators differ in their responses to environmental change, in this case wind speed. For the first time, we demonstrate an interaction between pollinators’ spatial foraging preferences and environmental change. At present, most response diversity functioning examples come from biomass and microcosms . Our results show that wild pollinators help to sustain pollination services under extreme weather conditions when the service by honey bees declines. We also show that functional pollinator taxa use different spatial niches depending on wind speeds. These results demonstrate complementarity among pollinators in foraging behavior and differential responses to altered environmental conditions,plastic pot manufacturers providing mechanistic support for a positive relationship between diversity and the magnitude of pollination services.

Diversity buffered almond trees from a reduction in flower visitation under altered environmental conditions. Individual almond orchards can be in bloom for up to 2 weeks. From 1983 to 2011, in February and March when the almonds are in flower, the number of days when the average wind speed exceeded 2.5 m s
1 was 13 and 15, respectively . There is therefore a high chance of high wind speeds during almond bloom. The differential abilities of wild bee species to fly and forage under high wind speeds differentiates their environmental niche from honey bees and our results show that their spatial niches can change under changing environmental conditions. Honey bees were the most sensitive visitor taxa to wind speed. The wild bee Andrena cerasifolii was particularly abundant at high wind speeds . The physiological and/or behavioral mechanism by which bees such as A. cerasifolii and O. cornuta can forage at higher wind speeds than honey bees are not well established and merit further investigation. A diverse community of flower visitors supplies a greater magnitude of pollination service in almondorchards by visiting a greater proportion of the spatial niches in the tree. Greater pollinator diversity has been associated with increased fruit set in almond . In Klein et al. , the fruit set in the different tree sections was not specifically compared. However, the present study shows that pollinator diversity can improve the spatial distribution of pollination service through complementary foraging locations within trees, providing evidence of a mechanism through which diversity can increase fruit set. Honey bees showed a preference for foraging in the top parts of the tree. Hover flies and other flies can be reasonably effective pollinators of almond flowers and they foraged more in the lower parts of the tree, filling the vacant niches. The foraging location of the wild pollinators may have been related to competition with the honey bees, different microclimatic preferences, and/or minimizing energy costs .

When fewer honey bees were present on sunny days with high wind speeds, the spatial preference of wild bees switched to the top interior of the tree, which under low wind speeds was heavily visited by honey bees. We did not observe many direct interactions between honey bees and wild pollinators suggesting that resource, as opposed to interference competition was more likely. Spatial complementarity can occur at different scales and has also been demonstrated within flowers in strawberries . At high wind speeds, honey bees preferred to forage in the bottom interior section of the trees. This section may be more sheltered from the wind, reducing the energy costs for flying between flowers and the oscillation of the flowers they are trying to land on . There can be differences between the shape of almond trees of different varieties, ages, and management. These differences may affect how exposed or attractive the different sections of the tree are to the different pollinator taxa. This study selected orchards with extremely diverse pollinator communities for comparison. For the many almond orchards isolated from natural habitat, management such as restoring flowering secondary habitat strips may be necessary to support wild pollinators . In addition, managed Osmia species can provide an opportunity for isolated orchards to diversify their pollination service. O. cornuta has been found to forage at lower temperatures and higher wind speeds than honey bees in apple orchards . With increasing demand for biotic pollination in crop production , the importance of wild insects and their functional diversity in sustaining pollination services and food production may become increasingly important over time and with environmental change. The presence of wild bees can help protect pollination services in the face of climate change by increasing the range of responses of pollinators to climatic fluctuations. In almond, under high wind speeds a dramatic drop in visitation by honey bees was buffered by wild bees . By only considering wind speed here, we may be underestimating the potential response diversity of pollinators . Following heavy rainfall, we noted that honey bees resumed foraging more quickly than wild bees . Other factors such as temperature and the timing of the onset of flowering may elicit different responses.

At present, there is limited knowledge of the diversity of response traits of wild bee and other pollinator species; however, this study demonstrates the importance of diversity for sustaining pollination services under just one of many potential changes in environmental conditions. Therefore, instead of relying on a single species, the conservation of pollinator biodiversity in general is recommended to help ensure the sustainability of pollination services in the future .Traumatic brain injury accounts for approximately 90% of brain injuries, and is associated with cognitive dysfunction and long-term disability.[1] As a result of domestic incidents, military combat, traffic accidents, and sports, TBI can compromise broad aspects of neuronal function. Patients often experience problems in the domains of learning, memory, and affective functions that can profoundly influence quality of life.Existing therapeutic strategies for TBI have not been successful in counteracting the heterogeneous TBI pathology nor improving the quality of life of patients.Hence, identifying interventions with broad applicability seems necessary for effective management of TBI. Dietary polyphenols have significant positive effects on brain health via protecting neurons against injury and enhancing neuronal function.Evidence supports the neuromodulatory effects of flavonoid-rich blueberry, particularly in promotion of brain plasticity,and counteracting behavioral deficits.In the United States, demand for blueberries has increased, with 2017 fresh per capita consumption of 1.79 pounds per person.Several reports indicate that blueberry dietary supplementation improves memory, learning, and general cognitive function,black plastic plant pots wholesale and protects against neuronal injury associated with stroke.Moreover, it has been shown that blueberries possess potent antioxidant capacity through their ability to reduce free radical formation or upregulating endogenous antioxidant defenses.These studies suggest that blueberry supplementation can have the potential to be used to overcome the broad pathology of TBI. Given the lack of information about the effects of blueberry intake immediately after TBI, we have performed studies to assess the effects of blueberry extracts during the acute phase of TBI. Evidence suggests that TBI is characterized by dysfunction in synaptic plasticity, elevated levels of free radicals, plasma membrane dysfunction,which can contribute to the behavioral dysfunction. Oxidative stress is part of the pathology of TBI and compromises neuronal function.In particular, excessive free radical formation leads to accumulation of lipid oxidation byproducts such as 4-hydroxynonenal with subsequent impairments in plasma membrane fluidity, receptor signaling across the membrane to deteriorate synaptic plasticity and reduce neuronal excitability.Deficiencies in brain derived neurotrophic factor reduce the brain plasticity necessary to cope with the effects of TBI.BDNF activates cAMP-responsive element-binding protein , a multifaceted transcriptional regulator involved in synaptic plasticity essential for learning and memory.BDNF is known to bind to TrkB receptors, leading to activation of Ca2+/calmodulin-dependent protein kinase II , required for synaptic processes involved in behavior.

Several observations indicate that the flavonoids exert action through modulation of signaling pathways to promote synaptic and neuronal function.Accordingly, in the current study, we investigated whether blueberry supplementation would counteract TBI pathology by involving BDNF-related pathways involved in synaptic plasticity and oxidative stress to influence cognitive behaviors.In the present study, we found that BB supplementation can attenuate important aspects of the acute TBI pathology. We report that BB supplementation immediately following TBI mitigates behavioral deficits in spatial learning and memory. BB supplementation counteracted the effects of TBI on proteins associated with the action of BDNF on plasticity and behavior. In addition, BB supplementation counteracted the increase of the end product of lipid peroxidation, 4-HNE. The results showing that markers of neuronal plasticity and lipid peroxidation change in proportion to memory performance suggest a possible association between these molecular parameters and behavior. Taken together, the present findings emphasize the beneficial effects of BB supplementation in fostering brain plasticity in the TBI pathology.In agreement with previous reports,we found that TBI impairs spatial learning as evidenced by an increase in latency in the Barnes maze, while BB supplementation decreased latency time to find the escape hole at each training day. We assessed the use of spatial learning strategies in our paradigm to provide a complementary measure of cognitive function less dependent on motor behavior. Interestingly, we found that BB supplementation appeared to counteract a lost capacity of TBI rats to employ spatial leaning cues. This information together with results of the shorter latencies strongly suggest that BB supplementation protects TBI animals from a loss in spatial learning performance. In this regard, recent functional neuroimaging study in humans has established a connection between BB intake and cognitive function.Further, in the EPM test, rats exposed to TBI showed a tendency to reduce time spent in the open arms, which encompasses with clinical reports that psychiatric disorders are often observed in TBI patients.TBI-induced behavioral deficits probably stems from the impairments in BDNFTrkB signaling that has been implicated in various cognitive and affective disorders.We cannot ascertain the cellular identity of the reported protein alterations. Although neuronal cells are the primary locus for learning and memory processing, nonneuronal cell types such as astrocytes and microglia can also contribute to these alterations.Moreover, it known that astrocytes and microglia provide support to synaptic transmission that is fundamental for neuronal function involved in cognitive processing.In the present investigation, we also found that TBI significantly reduced levels of hippocampal BDNF, and that BB dietary supplementation normalized these levels. Previous report indicated that deficiencies in BDNF signaling is associated with impairments in cognition.Alternatively, cognition is strongly reliant on long term potentiation and hippocampal BDNF, and the interaction between BDNF and its tyrosine kinase receptor is required for induction of LTP.Previously, we have shown the protective effects of BDNF on the TBI pathology.Presently, our findings show that BB supplementation counteracted the BDNF reduction induced by TBI, paralleling improvements in cognitive function. It is well established that BDNF regulates synaptic plasticity and learning through interaction with the transcription factor CREB.Interestingly, our results also showed that BB supplementation normalized levels of CREB in TBI animals, and that these changes were proportional to changes in BDNF levels. These findings are consistent with reports showing that BB dietary supplementation enhances BDNF-mediated plasticity with improved spatial and object recognition memory.Moreover, the significant positive correlation between levels of BDNF and CREB indicates that BDNF and CREB are co-regulated in our paradigm. In addition, evidence indicates an association between BDNF and CREB, and this interaction is important for regulation of learning and memory.The latter possibility can also be inferred from our results showing a negative correlation between CREB signaling and latency in the Barnes maze.

Fewer than half of California’s school districts participated in the program, but participating school districts had larger student enrollments than nonparticipating districts . The ethnic profile of students and the average school breakfast participation rates were similar in participating and nonparticipating school districts. School districts were stratified according to their number of elementary, middle and high schools, and the schools were randomly selected for participation in the evaluation. Of 93 schools that were contacted, 20 were ineligible because they were not participating in the program and four declined to participate in the evaluation. Of the remaining 69 schools, 61 were able to supply sufficiently complete data for the evaluation. The Committee for the Protection of Human Subjects at UC Berkeley approved the study. Parents received letters about the study, and students consented verbally to participate. Data were collected before and during the program. Breakfast menu production records and invoices were sought from the schools’ child nutrition directors on 20 randomly selected days during the months of September, October and November in the year before the program was implemented and during the program . Data from the menu production records included the nature and number of fruit servings prepared and taken by students at breakfast. Nonfood expenses directly relating to operating the program were also reported. Of the 61 nutrition directors, 55 recorded their views of the program’s impact on nutrition services operations, perceived student satisfaction, challenges and barriers to operating the program, nutrition education and promotional techniques, and staff training and needs. A stratified random sample of 18 schools was selected for site visits,raspberry grow in pots which were successfully conducted at 16 schools: six elementary, six middle and four high schools.

This sample was similar to other schools participating in the program in terms of school level, enrollment, geographic location, free- and reducedprice enrollment and student ethnicity. Interviews with nutrition directors were conducted at each of the 16 schools. Student surveys were completed by 1,205 students in grades 4 to 12 in a convenience sample of one or two classes at each of the 16 visited schools as well as at the school cafeterias during breakfast service. Questions were asked about where breakfast is eaten, how often fruits and vegetables are consumed at breakfast, favorite fruits to eat at breakfast, importance of eating fruits and vegetables at breakfast, change in fruit and vegetable consumption compared to the previous year and basic sociodemographic information. The cafeteria questionnaire asked additional questions regarding opinions about the school breakfast and perceptions of change since the previous year. The classroom questionnaire included questions regarding barriers to eating the school breakfast. In addition, trained research staff facilitated classroom discussions with students in 28 classes in grades 4 to 12 . Students were questioned about their views on breakfast in general, the School Breakfast Program, the California Fresh Start Program and factors influencing their school breakfast participation and food choices. Nutrition directors recorded School Breakfast Program participation on a standardized form. The researchers obtained monthly participation data during the course of the evaluation, including number of operating days and school average daily attendance. In addition, observations of the breakfast environment were made at each of the visited schools. Costs of specific fruits and vegetables were calculated from invoices provided by the nutrition directors. The costs of fruits and vegetables prepared and served were based on the total value of the prepared items reported on the menu production records. Nonfood expenses identified on invoices were classified as transportation, facilities, large and small equipment, material, promotional, training, additional staff time, and other. The percentage of total nonfood expenses for each category was calculated. Differences in both fresh fruit and total fruit taken by students and in the variety of fruits offered at each school were calculated from menu production records and analyzed by t-test.

Descriptive findings were reported for schools demonstrating more successful program implementation, specifically, schools with increases of 0.10 or more units of total and fresh fruit taken and increases greater than 0.90 for number of different fresh fruits offered. Although the California Fresh Start Program was designed to increase fruit and vegetable consumption, its effect was almost completely seen on fruit consumption, since vegetables were rarely included in the breakfast menu; vegetables represented less than 1% of produce offered to students. Thus the results presented here are based on fruit offerings. The California Fresh Start Program resulted in substantial increases in the variety of all, and especially fresh, fruits offered to students. More than twice as many different fresh fruits were offered per day during the program compared with the same period a year before: an average of 1.38 fruits compared to 0.66 . When considering all forms of fruit , there was a 46% increase in the average number of fruits offered per day: 2.55 fruits compared with 1.75 prior to the program . The California Fresh Start Program brought the offerings into compliance with dietary recommendations for two produce servings at breakfast. During the program, fresh fruit made up the majority of the fruit offered at breakfast. Juice, which previously had been the primary source of fruit, decreased substantially as a proportion of total fruit. All types of fresh fruit were offered with greater frequency; however, apples remained the most commonly offered individual fruit, followed by oranges and bananas. Stone fruits, though offered less frequently, showed the greatest percentage increase during the program . Our findings suggest that when offered a greater variety of fruits and less juice, students will increase their intake of fruit, especially fresh varieties . Students eating the school breakfast took more than twice as many fresh fruit servings during the California Fresh Start Program than before the program, 0.32 servings compared to 0.14, while taking substantially less juice and nearly the same amounts of canned, frozen and dried fruit offerings .

Although there were no direct measures of student consumption in this evaluation, the amounts taken, as recorded by food service personnel, provide a reasonable indirect basis for assessing student consumption. Observations by research staff and food service personnel confirmed that most students who choose to take a fruit at breakfast do eat it. Therefore, student consumption of fresh fruit at breakfast appears to have doubled as a result of the California Fresh Start Program. During the program, students took more of almost all types of fruit; however, the percentage increases were greatest for less common fruits such as cantaloupe, tangerines/tangelos and blueberries, which were not often offered before the program. Increases of about 20% to 30% were observed for common fruits such as apples, bananas and oranges; increases were 100% or more for tangerines, berries and cantaloupe, reflecting their appeal among students, and the low frequency with which they were offered before the program. Although the greatest increase in offerings occurred for the most common fruits — apples, oranges and bananas — the relative increase in servings of fruits was highest for the less common fruits. The demand for more common fruits may be approaching saturation,30 planter pot but unmet demand exists for a wider variety of fruits. Thus, future increases in the fruit servings students take at breakfast will likely require offerings of fruits other than apples, oranges and bananas.While the overall impact of the program on the amount of fruit — particularly the amount of fresh fruit — taken by students is impressive, this impact is even more dramatic when looking specifically at the schools that experienced the greatest success in implementing the program. At these schools, the California Fresh Start Program led to a 46% increase in the total amount of fruits taken by students, and a 383% increase in the fresh fruits and vegetables taken . It had the most impact in schools where students took the lowest number of fruit servings before the program — schools with the greatest need for an increase in produce intake. Schools that offered increased quantities of fruit, more variety of fruits and more unusual fruits and less juice were most successful in increasing student selection of fresh fruit. Limiting juice and providing fruits other than apples, oranges and bananas appear to be particularly important for increasing student consumption of fresh fruit. The fruits most often served at breakfast are rarely the ones that students most prefer .Students’ attitudes toward eating fruit, already positive, showed modest changes during the program. Most students reported it was important to have fruit at breakfast, saying that fruits and vegetables are “good for you because it’s healthy, makes you strong; there is natural sugar, and it contains vitamins like A and C.” However, only 13% said they always eat fruits and vegetables at breakfast, and only 19% said they often do. This maybe due in part to the fruits most often served at breakfast not being the fruits students prefer. Students prefer more exotic fruits than they are currently served. Fruits mentioned were mangos, kiwi, strawberries, peaches, pineapple, watermelon and grapes, with melons and berries being most popular . Students also want more variety in the ways fruit is presented, including chopped fruit, fruit salads, salad-type fruit bars, fruit with condiments and ethnic favorites. Variety, convenience, quality and freshness are key concerns. High school students, in particular, expressed a desire for more tropical fruits such as mangos. Successful implementation of the California Fresh Start Program was not significantly related to school characteristics or student socio-demographics. The type of school , ethnicity/race of the students, ruralurban geographic location, percentage of free- and reduced-price meal participation and size of student enrollment did not have any statistically significant association with the program’s success in terms of the number of fresh and total fruit servings students took or the variety of fruits offered. Whether students were offered and/ or took more fruit was affected by a variety of school institutional and economic factors, including the physical layout of the school’s food service department and the availability of funding for program support.

Because the California Fresh Start Program did not mandate or provide funding for facility improvements, it is not surprising that only about 9% of the schools made improvements to their kitchen, dining area, serving areas or points of service. Data from the evaluation indicated that adequate dining space for students was related to students taking more fruit and an appealing dining ambience was related to students taking more fresh fruit. Almost one-third of schools did not have sufficient facilities to seat all students comfortably. The temperature was uncomfortably cold in many of the serving and dining areas, which may have played a role on cold days in students’ preference for hot breakfast items rather than cold fruit. The student survey revealed that a majority of students want more options regarding when and where they can eat breakfast, particularly the options of eating in the classroom and indoors or outdoors. Only about one-third of schools offered students the choice of eating indoors or outdoors. Our findings suggest an investment in facilities has the potential to attract higher participation in the breakfast program and to increase students’ intake of fresh fruit. Schools that offered more variety of fruits were more likely to have made improvements in customer service, nutrition education, student attitudes and the quality and appeal of the fruit offered. Quality concerns were prominent in discussions with students about the changes in foods offered. Students noticed both positive and negative changes in food and beverage temperatures, freshness, taste, portion size and preparation. In addition to presentation, the position of fruit in the serving sequence might affect student selections. At one site, the fruit was not visible; it had to be requested. Fresh fruit was the first item offered in the serving sequence at only three of the 13 sites where these data were recorded. Nutrition directors at schools where students chose more fresh and total fruit were more likely to describe inadequate storage space and facilities. Many schools increased nutrition education and promotion efforts among students as part of the California Fresh Start Program but lacked the staff time and resources to mount a sufficiently intensive effort. Of the nutrition directors surveyed, 96% reported that lack of opportunity was a barrier to fully providing the nutrition education component of the program, 87% reported a lack of staff time as a reason, and 81% reported that lack of funding was a barrier .

If PCI reaches even 0.5% of PLW, it could cost USD $144 M. Additional costs related to 1) shipping at temperatures higher than the commonly used 4°C, and 2) the complex logistics that factor in harvest date and storage life into transportation, are not included into this estimate, but they collectively reduce flexibility for growers and distributors. Periodically, the apparent benefits of extending shelf-life by storing produce at inappropriate temperatures may outweigh the negative impact on quality: if fruit are stored at non-PCI-inducing temperatures for the equivalent time, they may spoil and will be rejected, whereas fruit with invisible PCI symptoms are salable. This incentivizes refrigerating sensitive produce, which may be profitable in the short term, but leads to long-term consumer distrust in produce quality and value. The abnormalities associated with PCI that lead to consumer dissatisfaction, waste, and loss can be linked to specific cellular dysfunctions . Mealy texture, surface pitting, and fungal susceptibility are due to reduced pectin solubilization and depolymerization, and microfractures in the cell-wall network . Tissue browning is initiated when organelles lose their structural integrity in chilled tissues. Chilling leads to membrane disassociation that releases polyphenol oxidase, peroxidase, and their phenolic substrates into the cytosol where they react to form quinones. Polymerization of quinone creates the brown pigments in chilling-injured tissues. Cold storage disrupts the finely-tuned ripening program that is modulated by the interplay of hormones,blueberry in container transcriptional factors , and epigenetic marks often, with negative consequences for fruit quality.

Upon rewarming, increases in respiration and ethylene production are initiated, and visible chilling-injury symptoms develop, the magnitude of which is inversely proportional to the storage temperature. Even mild chilling injury in red tomato fruit triggers epigenetic changes in ripening TFs such as RIN, NOR, and CNR that down regulate the production of key volatiles responsible for hedonistic values . However, this is not always so, ‘abnormal chilling injury behavior’ occurs at milder storage temperatures and has been reported in peach and nectarine, plum, persimmon, and papaya. The mechanisms underlying ‘abnormal chilling injury’ are unknown, but the enhancement of sugar and energy metabolism may be relevant. Many economically important commodities, for example, zucchini, cucumber, and bell pepper, are harvested before reaching physiological maturity, and will thus have different cold-stress responses to those commonly studied such as tomato and banana, which are harvested when mature. Commodities harvested at immaturity typically have higher respiratory and deterioration rates, greater water loss because of incomplete cuticle development, and inefficient reactive oxygen species scavenging systems, which will influence their PCI response. In potato, chilling leads to cold-induced sweetening — starch breakdown and sugar accumulation, where the latter serves as protective compatible solutes. When these ‘sugared’ tubers are fried, baked, or roasted, they turn black as acrylamide forms, which is visually unappealing, bitter in taste, and harmful to human health. Reducing the severity of the negative traits of PCI that lead to waste and loss could be achieved by inducing allelic diversity at single or multiple genes that directly regulate relevant pathways . Integration of multi-omics data of cold-injured tissue compared with non-chilled tissues, has helped identify gene targets that influence PCI.

In tomato, two important discoveries were made from genes identified using functional genomics: over expressing the TF SlGRAS4 reduced fruit-surface pitting, and promoted a more uniform color due to increased β– carotene content after chilling. Likewise, over expression of SlCYP90B3, a key brassinosteroid biosynthetic gene, improved the antioxidant response of fruits during cold storage, reducing PCI. These genes coordinated multiple pathways to improve PCI tolerance . Candidate genes for improving PCI tolerance have also been found by applying physical and chemical treatments that alleviate symptoms, and by studying the associated changes in the signal-transduction pathway. This is an active area of research where the literature is expanding rapidly. For example, physical treatments such as dipping in hot water before chilling mitigated fruit PCI, and have been associated with the upregulation of heat-shock proteins in banana and in mango, among others. The MaAPY gene family and the ATP receptor MaDORN1.19 are likely to be important for maintaining ATP homeostasis under chilling, after exogenous application of ATP or GTP to banana fruit. TFs MabHLH060 and MabHLH183, which are associated with reduced cold-induced membrane rigidity, were induced by Ethrel®, an ethylenereleasing agent, which also reduced banana fruit PCI visual symptoms. Melatonin reduced peel browning in bananas, by inducing miR528 expression, which in turn down regulated several ‘browning genes’, i.e., MaPPO1, MaPPO2, and MaPPO3 . In tomato fruit, melatonin improved surface pitting, increased the expression of FAD3 and 7, and reduced the expression of phospholipase Dand lipoxygenasegenes, which helped to maintain membrane integrity under cold stress. New insights into regulatory networks governing PCI can be gained through-omics profiling as shown in several examples. Treating peach fruit with methyljasmonate delayed internal browning , maintained fruit texture and aroma volatiles, and the accompanying transcriptomic and methylomic changes were revealed.

In bell pepper, MeJA reduced surface pitting, shriveling, discoloration, and seed browning, and differences in the transcriptome, proteome and metabolome compared with untreated fruit were detected. Other studies combined cold storage with additional factors such as varying carbon dioxide, or fruit harvested at different times, and identified differentially expressed transcripts by RNASeq. These differential transcripts, proteins, and metabolites may help to identify gene networks and their regulators for genetic engineering. Gene targets for minimizing CIS in potato have been identified. CIS has been extensively studied compared with PCI in fruit, because potato is a staple for one billion people, and in some production areas, tubers are stored at low temperature for up to eight months. During tuber storage, there are cycles of synthesis and degradation of both sucrose and starch, but at temperatures between 4 and 10°C, the degradative fluxes are activated, so that reducing sugars accumulate . This change in metabolism occurs via the upregulation of genes encoding the beta-amylase, glucan water dikinase, sucrose phosphate synthase, and invertase enzymes . Attempts to alleviate CIS by modulating the activity of core enzymes of carbohydrate metabolic pathways have been made, although the role of each enzyme isoform is still to be elucidated. There is an acute need for a greater foundational understanding of PCI. Several advances have been made in model species, where regulatory elements of the cold signal-transduction pathway response have been identified and functionally verified. Integrating the discrete ‘snapshot’ studies discussed previously into full models across tissues, developmental stages, and conditions, is the next step for developing functional biotechnological solutions. PCI is often assessed in a single tissue sampled from a defined region. Not only is valuable information about the spatial evolution of the process lost , but events occurring in all the tissues that are consumed are not captured. A few studies have addressed this gap and serve as a guideline for future work. In tomato, tissue specific development of PCI was detected, even though pericarp is usually the only tissue traditionally studied. In pineapple, scanning electron microscopy and histochemical staining of fruit revealed that IB starts at the phloem and diffuses throughout fruit tissues from the core. A breakthrough was made when a high resolution spatiotemporal transcriptome atlas in tomato was developed, plastic planters bulk which showed that ripening is not homogeneous. Because cold interferes with fruit ripening, some PCI symptoms would be expected to occur heterogeneously. Looking holistically at the chilling response across cells and tissues in harvested organs would uncover additional regulatory features of PCI. Uncontrolled and physiologically abnormal expression of genes through genetic engineering may severely disrupt the multiple finely-balanced gene-regulatory networks, resulting in deleterious phenotypes, especially if constitutively expressed in tissues where they do not normally occur. Regulated promoter systems to direct tissue gene expression in a highly controllable manner, with spatial and temporal precision, may be useful to study and design long-term solutions to PCI. Sequential changes in gene expression by promoter engineering are also a promising approach. Precise editing of chilling-associated cis-regulatory elements and differentially methylated regions due to chilling by Clustered Regularly Inter spaced Short Palindromic Repeats could promote ripening under chilling conditions and alleviate PCI .

Achieving this precision is an immensely challenging prospect that likely depends on attaining the previously described holistic knowledge of chilling response. There are many longstanding challenges related to PCI that remain largely to be tackled, and which limit progress. The nature of research appears fragmented, and many species are studied with a substantial focus on symptom alleviation through exogenous treatments rather than development of endogenous/innate solutions. The importance of PCI is reflected in the number of papers published in Google Scholar using the search term ‘Postharvest chilling injury’, which has increased 7- fold over the past 20 years . This directly points to the urgency of developing biotechnological approaches to address PCI, and the immediacy with which solutions are needed. Environmental and management factors, both pre- and post harvest, influence whether a tissue will manifest PCI after cold storage. Time of year, time of day of harvest, and the growing environment are all documented to influence the trait . Experiments are by necessity, multi-factorial, and rely on large harvests of fruit, tubers, and so on, which limit the number of experiments that can be set up. In addition, plants must reach advanced developmental stages to obtain fruit or tubers, and this is followed by weeks of post harvest cold storage, limiting the number of experiments that can be performed in a year. This is exacerbated in perennial crops that fruit annually and may be biennial, offering fewer opportunities for experimentation. Replication of experiments may not be economically feasible, therefore, at a minimum, a thorough reporting of these factors should be included in PCI studies. There is a limit to which the data so derived can be translated to PCI. The growing plant may use avoidance, escape, and tolerance to cope with cold, while in contrast, options for surviving anthropogenic cold stress in stored tissues with limited nutrients are few . Coldresponsive genes in Arabidopsis rosettes may have conserved functions in leafy greens, but genes and pathways from model crops, for example, cereals, will have limited relevance to the consumed tissues in horticultural crops. Therefore, post harvest studies must redescribe the behavior of these pathways under the conditions of each experiment, which is laborious and expensive or work with tempered assumptions about them.The question of if there is a single primary event that triggers others, or if multiple events occur simultaneously, remains. ROS production, and membrane disassembly are cited as incipient processes, but their relative timing has not been resolved, as there are contrasting reports of their relative importance. Studying biological process hours after chilling would illuminate the rapid and early, cold-tolerant responses that are often overlooked. Understanding the progression of these events in different tissues could allow for more targeted and efficient solutions in alignment with the spatial and temporal occurrence of PCI events. Molecular components of the chilling pathways in tomato, peach, banana, and potato have been identified because of their economic importance and advanced functional genomics tools, while research on other species lag. However, whole genome sequencing of diverse crops such as pineapple, coconut, and basil is fueling PCI research in these species. Bridging the technological gap for PCI research in tropical species would benefit multiple stakeholders, astropical commodities are rich in bio-active compounds and have exotic tastes and flavors, of interest to international markets. These crops are likely to suffer from PCI that increases export costs, since air transport, rather than maritime shipping, would be needed. The importance of day length for plant photosynthetic carbon gain has been widely recognized. Duration of the nocturnal period may also be important. During the nighttime, periods of minimal transpiration allow water potential gradients among plant parts to dissipate by internally redistributing water to tissues of lower water potential, including the movement of water to roots in dry soil layers . Nocturnal internal hydraulic redistribution may be very important to the maintenance of roots in surface soil layers where root densities are highest, as surface soil can be dry for extended periods in many regions . However, the rate at which water redistributes at night depends on a combination of environmental and plant factors, including the magnitude of differences in water potential among tissues and the hydraulic conductivity of the vascular system . Many factors influence the rates of tissue rehydration of roots at night.

The total tonnage of fertilizer applied to corn increases by 484.45 tons, an 26.6 percent increase, statistically significant at the 1 percent level. The impact congregational mergers have on corn fertilizer use is not only statistically significant, but also represents an economically meaningful change. In sum, the information in Table 2.3, Table 2.4, and Table 2.5, demonstrates that congregational mergers increase fertilizer use, in keeping with Proposition 2. Next, we test Proposition 3: that congregational mergers will not affect technology adoption when all potential adopters are fully informed. Strip cropping, irrigation, and orchards are technologies for which we expect no effect. Table 2.6 demonstrates the impact congregational mergers have on the number of farms using strip cropping and the acres under strip cropping; the number of farms reporting irrigation use and the total number of irrigated acres; and the total number of acres in fruit orchards, groves, vineyards, and nut trees. Table 2.6 confirms our hypothesis: we see no statistically significant impacts of congrega- tional mergers on strip cropping, irrigation, or orchard acreage. Furthermore, the magnitudes of the coefficients we do see are small, and in the case of irrigation, have negative signs. The absence of results in this table further supports the fact that congregational mergers are driving the changes in input uses observed above, because mergers are affecting farmers on the dimensions we would expect, but we are not seeing impacts on agriculture that should not be affected by mergers.With any regression analysis,blueberries in containers growing it is important to ensure that the results are robust. We use three pieces of evidence to demonstrate that the results we present in this paper hold up to robustness checks. First, we saw impacts of congregational mergers where we expected them: on fertilizer and lime use and on corn. We did not see impacts where we expected them not to be: on strip cropping, irrigation, and orchards, as presented above.

In addition, our results are robust to a variety of different land use variables provided in the census of agriculture data: the outcomes measured in number of farms do not change significantly if instead of total farms, we use commercial farms or cash-grain farms; the outcomes measured in acreage and tonnage do not change if we use acres in the county or acres harvested rather than acres in farms. In this section of the paper, we perform a placebo test. We also investigate channels other than information through which congregational mergers might be driving fertilizer adoption, and provide evidence against these other possible explanations.Table 2.7 has the placebo analysis. We run this check in order to see whether mergers that occurred from 1964 to 1967 impact our outcomes in 1964, before these mergers actually occurred. We use the time period 1964 to 1967 because it includes the same number of years as our actual treatment period. Column looks at farms using fertilizer; at acres fertilized; at tons of fertilizer used; at corn acres fertilized; at farms using lime; and at acres limed. We expect to see no statistically significant impacts of “future” congregational mergers on these outcomes. Indeed, Table 2.7 shows that, with the exception of the number of farms using lime, there is no statistically significant effect of future congregational mergers on 1964 input outcomes. In addition, comparing these effects to those in Tables 2.3, 2.4, and 2.5, the magnitudes of the coefficients are quite small. This helps confirm that the effects we are observing above are real and driven by the congregational mergers we observe, rather than by something unobserved. To assuage concerns about small-sample inference, we also run a permutation test, in which we randomly assign 39 counties to treatment 10,000 times. Using each of these treatment assignment vectors as a placebo treatment, we find our actual effect on farms using fertilizer is larger than all but 4 percent of the randomly drawn treatment vectors. Figure 2.7 displays the results of this procedure. It is still possible that our results are being driven by something other than a congregational merger driven information effect. Here, we explore two other possible explanations for our results. The first is the presence of agricultural extension.

Agricultural extension, formally introduced in the United States by the Smith-Lever Act of 1914, plays a major role in information dissemination in agriculture. There is a large literature on the effect of agricultural extension, both in the United States and elsewhere, on agricultural productivity and technology adoption ; Huffman ; Birkhaeuser et al. ; Dercon et al.. Despite the importance of extension, we argue that it is in fact congregational mergers and not extension services that generate the results we find in this paper: because of the fixed effects strategy, in order for agricultural extension to be driving these results, we would need to see agricultural extension services changing differently over time in treatment counties than in control counties, having removed the state time trend, only over the 1959 to 1964 time period. This is potentially plausible, but seems unlikely, especially because extension funding and the number of extension agents allowed is governed by state laws, which do not change often. For example, the Minnesota statutes outlining extension were first passed in 1923, updated in 1953, and were not revised again until 1969. The law allows for “the formation of one county corporation in each county in [Minnesota]” to act as an extension agency, with in most cases one extension agent and a specified budget, based on the number of townships in the county. While county extension offices documented their activities for mandatory state reports, these reports were inconsistent across different counties and years. Also, many of the variables measured were endogenous, such as the number of phone calls received or the number of attendees at extension events. As a result, it is impossible to credibly measure the intensity and efficacy of extension efforts over our sample period.Another plausible explanation would be that the mergers also facilitated increased access to capital. In order to provide evidence against this possibility, we estimate Equation again, this time with the number of farms with each of a variety of capital-intensive technologies as outcome variables. Table 2.8 shows the impact congregational mergers have on the number of farms with cars, trucks, tractors, bailers, and freezers.

As expected, we find no statistically or economically significant effect of congregational mergers on capital-intensive inputs: the standard errors are quite wide, and the effect sizes small: the coefficient on cars, for example, is only a 0.01 percent increase relative to the control group mean, and the standard error is almost one hundred times the size of the coefficient. This suggests that congregational mergers did not substantially increase access to capital, and provides additional evidence that information is the main channel through which congregational mergers impacted technology adoption. Finally, one might worry that by only using TALC congregational mergers in our analysis, we are understating the true treatment effect. We argue above that the TALC mergers are exogenous, and, due to the heavily Lutheran populations in these regions, the mergers where we would expect to see an effect. Indeed, the congregations that are merging in these data have, on average, 492 baptized members, so seeing an additional 35 farms begin to use fertilizer is an entirely reasonable effect size. There is another major Lutheran church branch, the Lutheran Church – Missouri Synod ,planting blueberries in containers that was not directly involved in the TALC merger, but whose mergers could be attributed to increased discussion about merger surrounding TALC. We collected data from Concordia Historical Institute, the LCMS seminary, on congregational mergers between LCMS churches during the sample period. There is only one merger that occurs in a non-metropolitan county during this time period, and the inclusion of said merger does not produce a statistically distinguishable result from using only the TALC mergers. Ultimately, given the range of tests that we perform, we have confidence that our results are robust and that we are correctly attributing them to the information effect of congregational mergers. Since the early 2000s, US ethanol production has exploded in response to federal policies incentivizing the production of renewable fuels. In 2005, Congress passed the Energy Policy Act introducing a Renewable Fuel Standard mandating that 2.78% of gasoline sold in the US be from renewable sources. In 2007, Congress passed the Energy Independence and Security Act setting annual renewable fuel mandates for US production with an ultimate goal of 36 billion gallons by 2022. Of these 36 billion gallons, 15 billion are to be conventional bio-fuels – corn-based ethanol in particular. The US ethanol industry has clearly responded to the Renewable Fuel Standards established in the EPAct and EISA. Between 2002 and 2014, US ethanol production has increased from just over 2 billion gallons per year to over 14 billion gallons per year . In order to produce such quantities of ethanol, the number of corn ethanol refineries in the US has increased from 62 in 2002 to 204 in 2014 . The striking increase in US corn ethanol production has raised several important questions about its unintended consequences. One strand of research has explored how increased demand for ethanol has affected land use in the US corn belt as aggregate demand for corn increases . Another strand of research has been more concerned about the environmental externalities of changing agricultural patterns, particularly focused on nitrate runoff and water pollution . In this chapter, I explore both the land use change effects and environmental effects of expanding ethanol production. In particular, I study the geospatial effect of ethanol re- fineries’ placement on nearby land use change and use my results to estimate environmental consequences. I am specifically interested in how the location of ethanol refineries spatially affects agricultural land, and I do not attempt to identify the full general equilibrium effect of the 14 billion gallon US corn ethanol industry. Put another way, I study how the distribution of ethanol refineries differentially affects different agricultural areas net of the ethanol industry’s aggregate effect on corn prices. I find that within a population of almost 114 million acres of agricultural land in Illinois, Indiana, Iowa, and Nebraska, nearly 300,000 more acres of corn were grown in 2014 than in 2002 due merely to ethanol refinery location effects.

This represents approximately 21,000 tons of nitrogen applied as fertilizer. Almost all the 300,000 acres of increased corn acreage exist within 30 miles of an ethanol refinery, suggesting that these refineries have strong local effects on land use change and nitrogen use. There is clear economic intuition for why ethanol refineries would differentially affect nearby and faraway agricultural land. When a corn-fed ethanol refinery is built, it represents a new terminal market for corn. Since refineries operate continuously, they have an inelastic demand for this input. And since transportation costs are significant for grains, one would expect an ethanol refinery to source its corn from the nearest producers. Thus, by reducing transportation costs for nearby producers , ethanol refineries essentially subsidize corn production for nearby farmers. On the margin, this subsidy incentivizes farmers to grow more corn – or grow corn more often – than they otherwise would. As corn production increases, so will nitrogen fertilizer use. Corn requires higher levels of nitrogen fertilizer than other Corn Belt crops, and particularly high levels of fertilizer when grown successively corn-after-corn. Thus, economic intuition suggests ethanol refineries would have a localized effect increasing corn production and nitrogen fertilizer use. Consequently, these refineries would also have an effect on localized nitrate runoff due to the increased nitrogen fertilizer use. Researchers have previously addressed different components of the ethanol industry’s effects on land use change and nitrate runoff. One line of research has explored whether the hypothesized local corn subsidy provided by nearby ethanol refineries actually exists. In a frequently cited paper, McNew and Griffith find that corn prices at an ethanol refinery are 12.5¢ higher than average, that the effect is slightly stronger for “upstream” refineries than for “downstream” refineries, and that price effects can be detected up to 68 miles from a refinery. However, Katchova and O’Brien both fail to find such a subsidy. Gallagher et al. highlight that locally-owned and non-locally-owned refineries have different effects on corn prices: the authors find that corn prices are increased by proximity to a non-locally-owned refinery, but not by proximity to a locally-owned refinery. Finally, Lewis finds different results in different states: ethanol refineries in Michigan and Kansas affect local corn prices, but refineries in Iowa and Indiana do not.

A study conducted on diabetic Zucker rats supplemented with an anthocyanin-rich extract from purple potato and bilberry, showed a reduction in dysbiosis of colonic microbiota and anthocyanins gut microbiota-derived metabolites, increased cecal sugar levels together with the increase in the abundance of Peptostreptococcaceae sp. and Parabacteroides spp. in colon. These results demonstrate that anthocyanins by modulating the gut microbiota can affect gut function and consequently prevent T2DM . Our RNAseq analysis showed for the first time that an anthocyanin-rich BB extract can affect the expression of large number of genes in adipose tissue, not only protein coding but also non-coding genes like miRNAs or lncRNAs. According to our knowledge, very few studies have reported the multigenomic effect of BB anthocyanins, as well as isolated anthocyanins, on adipose tissue. It has been observed that cyanidin 3-glucoside, anthocyanin from BB, can up-regulate the expression of GLUT4 gene in vitro in 3T3-L1 cells . The BB extract rich in flavonoids and saponins was also shown to exert the capacity to modulate the expression of key genes involved in lipogenesis, such as SREBP1c, ABCG5, CPT1, SREBP2, FAS, HMGCR, INSIG1 and INSIG2 . Supplementation of mice diet with BB has been reported to modulate the expression of the SCFAG-protein coupled receptors in colonic tissue as well as the expression of genes regulated the integrity of the epithelial barrier: occludin, JAM-A, E-cadherin and the junction/cytoskeletal connector ZO-1 . Recently it has been observed that black bean concentrate improves hepatic steatosis by reducing lipogenesis and increasing fatty acid oxidation in rats fed a high fat-sucrose diet . However, these studies present an important drawback,large plastic pots for plants which is use of targeted approach to assess the expression of few specific genes.

Rare recent studies have shown that anthocyanins can affect large number of genes presenting multi-modal action. For example, anthocyanins from blackberries affected the expression of over 600 genes, including miRNAs, in circulating immune cells in human volunteers . Another study has demonstrated the capacity of anthocyanin-rich bilberry extract to modulate the expression of over 1,600 genes in hippocampus of ApoE-/- mice . Enrichment pathway analysis in adipose tissue were performed using 406 differentially expressed genes following BB extract intake. This allowed us to show that the black bean extract can affect genes regulating cell signaling pathways in adipose tissue from diabetic rats, pathways related with phosphatase activity, phospholipid metabolism, phosphatidyl inositol 3-kinase binding, phosphatidylserine binding, and ER organization. On the other hand, we also found down-regulated pathways that participate in the modulation of NIK/NF-κB signaling, regulation of response to extracellular stimulus, positive regulation of cell junction assembly, negative regulation of cell population proliferation and negative regulation of cytokine-mediated signaling pathway. Remarkably, while regulation of NIK/NF-κB signaling is up-regulated in the DB group, we observed that the treatment with BB extract down-regulated this pathway in the BB group, suggesting the capacity of the extract to counteract the effect of T2D. The biological relevance of this pathway on adipose tissue is the critical function in reprogramming the fat cell transcriptome to inflammation in response to over nutrition and metabolic stress. The inhibition of NF-κB signaling has metabolic advantages for adipose tissue inflammation caused by obesity . On the other hand, NF-κB inducing kinase is a key controller of immunity and inflammation at local and systemic levels on metabolic processes.

In adipose tissue, NIK promotes adipogenesis by activating non-canonical NF-κB pathway, demonstrated by Pflug et al., when NIK deficient mice fed with a high fat diet showed decreased overall fat mass, increased insulin sensitivity, and energy expenditure . Moreover, anti-inflammatory effects have also been attributed to anthocyanins from Hibiscus syriacus L. by the capacity to inhibit TLR4 in LPS-induced cells, followed by the decrease of the phosphorylation of MyD88 and IRAK4, which resulted in NF-κB inactivation . Black soybeans also demonstrated to inhibit IκB phosphorylation that impede NF-κB translocation resulting in the inhibition of iNOS transcription and iNOS and COX-2 translation . Taken together, these results suggest that extract from BB can exert health properties by countering the genomic modifications induced by T2DM. Elevated levels of FFAs, inflammation, excess of nutrients, inadequately folded proteins, and local hypoxia, are characteristics of obesity and can lead to ER stress. This results in increased oxidative stress in adipose tissue of obese animals . ER plays an important role in adipose tissue inflammation due to their participation in the activation of NF-κB signaling by the phosphorylation of JNK, IKK, and JNK-mediated phosphorylation of IRS1/2 triggering the unfolded protein reaction and involving pathways like PERK , IRE1 , and ATF6 . Exposure of J774A.1 macrophages to a cyanidin-3-O-galactoside-rich aqueous extract of Sambucus ebulus L. resulted in suppression of LPS-induced transcription of pro-inflammatory genes like IL-1β, IL-6, TNF-α, Ccl2, Icam-1, Fabp4, COX2, iNOS, Noxo1, IL-1ra and Sirt1. SE also showed to produce the decrease in protein levels of iNOS, peIFα, ATFα and C/EBP Homologous Protein . Similarly, Punica granatum L. a high-content polyphenols Chinese plant, was administrated to diabetic rats for 4 weeks. It was observed that doses between 50 and 100 mg/kg can improve ER stress signals including IRE1, activation of XBP-1, as well as lower levels of IREα, XBPs, and CHOP . Therefore, anthocyanins present in our extract also have the capacity to modulate the expression of genes regulating ER stress, presenting another important molecular target underlying the anti-diabetic effects.

Our bio-informatic analysis also revealed that phosphatidylinositol 3-kinase binding signaling pathway was up-regulated in BB group. The importance of this pathway in adipose tissue is due to its role in adipose hypertrophy which rises tissue immune cell infiltration, fibrosis, and lipolysis. Decreases IRS-1 activation and AKT-induced glucose uptake, as well as exacerbates systemic insulin resistance and the development of T2DM. Different factors like adipocytokines and adipose hypertrophy produce insulin resistance by obstructing PI3K/AKT-mediated inhibition of lipolysis reducing the capacity of glucose utilization and diminishing the capacity of SREBP to stimulate lipid synthesis . Recent research concluded that sweet potato leaf polyphenols might up-regulate the important mediators of the insulin-mediated PI3K/AKT/GSK-3β signaling pathway in a dose-dependent way in diabetic mice by regulating the mRNA expression of genes IR, IRS-1, PI3K, AKT and GLUT-4 . Bioinformatic analysis of differentially expressed genes permitted to identify potential transcription factors whose activity can be affected by BB extract and result in the detected nutrigenomic modifications. Among these transcription factors are GATA2, POU2AF1, IRF3, GATA1, NR2F2 or PPARA. Interestingly some of these transcription factors have been identified as playing a role in adipose tissue development and/or diabetes. For example, it has been shown that GATA2 plays an important role in diabetes development and associated diseases . It has been described that hyperinsulinemia, observed during Type 2 diabetes, can activate NR2F2 which can induce development of different diseases . PPARs have central role in lipid and glucose homeostasis, pathogenesis of insulin resistance and metabolic syndromes and are active in adipose tissue. It has been shown that binding phenolic and other molecules to PPARs result in significant changes in their activities and present important mode of treatment type 2 diabetes mellitus. . Similarly, IRF3 has been described as a main transcriptional regulator of adipose tissue inflammation and is involved in preserving systemic glucose and energy homeostasis . One study also observed that IRF3 in adipose tissue promotes adipose inflammation and insulin resistance . These observations from our bioinformatic analysis reveal significant regulators of BB extract underlying their observed health properties. Moreover, our genomic analysis revealed that black bean extract rich in anthocyanins can also modulate the expression of microRNAs. MicroRNAs are short, non-coding RNAs which can bind to mRNAs,plant pots with drainage resulting in modification in translational repression. It is estimated that there are approximately 2200 miRNA genes in the human genome that can regulate the transcriptional levels of over 60% of genes. Consequently, they can regulate many major cellular functions such as development, differentiation, growth, or metabolism . Moreover, it has been shown that modifications in their expression produce an important role in progress of diseases. Among these, it has been demonstrated that miRNA might have an important role in development of diabetes and metabolic disorders . On the other hand, several previous studies have shown that anthocyanins can modulate the expression of these non-coding RNAs. For example, it was shown that a mixture of anthocyanins as well as their gut microbiome derived metabolites at physiologically relevant concentrations can affect the expression of miRNAs in isolated human primary endothelial cells . Also, a study showed that supplementation of mice diets with different polyphenols at nutritionally relevant doses can significantly affect global miRNA expression profile in liver . Among the miRNAs identified as differentially expressed by black bean extract are miR-152, miR-219a1, miR-384 or miR-615.

It has been observed that miR-152 is expressed in adipocytes and can stimulate lipid accumulation in preadipocytes accompanied by higher expression of some pro-adipogenic genes, adipogenesis and intramuscular fat formation . Its expression is also altered in patients with T2D with or without medication . miR-219a1 has been suggested to show a role in fat development. Similarly, it has been demonstrated that miR-615 is expressed in adipose tissue in humans and that its expression changes with the obesity . Moreover, the expression of this miRNA has also been identified in patients with diabetes . Together with miRNAs, our analysis also revealed changes in the expression of long non-coding RNAs. Biogenesis of lncRNAs is different than mRNAs and can interact with DNA, RNA, and proteins, modulating chromatin function, changing the stability and translation of cytoplasmic mRNAs and interfering with signaling pathways . Because of their large mode of action, they affect numerous cellular and consequently physio-pathological processes. It has been shown that they regulate adipogenesis and adipose tissue function and diabetes . Only few studies have suggested that polyphenols are potent modulators of the expression of noncoding RNAs. For example, epicatechin metabolites can regulate the expression of these miRNAs in human brain endothelial cells in physiologically relevant conditions . Therefore, our study provides novel and original data about the capacity of legumes and their bio-actives to affect the expression of this class of RNAs. However due to the lack of known biological functions of these lncRNAs as well as their targets, regulated cellular and molecular processes remain to be identified. Taken together, our results suggest that black bean extract rich in anthocyanins exert protective properties by modulating the expression of miRNAs and lncRNAs in adipose tissue in diabetic conditions.From a microbiological perspective, soil is largely unexplored even though we know it has a rich diversity of microbial life. Depending on its physical and chemical properties, soil can contain 109-1010 microbial cells per gram, including tens of thousands of different bacterial, archaeal, and fungal species, plus viruses and protists. Soil microbes carry out life-sustaining functions for our planet, including cycling of nutrients and promoting plant growth. Respiring soil microorganisms, producing metabolic byproducts such as carbon dioxide and methane, cycle enormous volumes of carbon-containing gases from the terrestrial ecosystem into the atmosphere. Perturbing this ecosystem—for example, when global warming raises temperatures—potentially alters the flux of these gases. Despite the immensity of the carbon reservoir in soil , its fate in the face of climate change is not known. The high diversity of soil microbial communities makes them difficult to study. Exacerbating this difficulty, few soil microorganisms are amenable to isolation and cultivation, steps that in conventional terms are crucial for elucidating microbial physiology and biochemistry. Although most isolated soil bacteria are “weeds” such as pseudomonads and actinobacteria, 16S rRNA gene surveys reveal that few species from the great bulk of soil bacteria have cultured representatives. For example, members of the Acidobacteria phylum are widespread in soil but are notoriously difficult to cultivate. To address these issues, microbial ecologists are now using a metagenomics approach, sequencing soil DNA to develop a better understanding of the microbial identities and their potential functions in soils. Initially, investigators who used metagenomics to study soil microbial communities cloned DNA extracted from soil into bacterial artificial chromosome vectors or fosmid vectors. One advantage of this cloning-based approach is that it permits investigators to screen for expression of particular phenotypes, including production of antibiotics and enzymes.