Phenotypic and molecular characterization of clinically isolated antibiotics-resistant S. aureus (MRSA), E. coli (ESBL) and Acinetobacter 1379 bacterial strains.

Antibiotic-resistant bacteria causing nosocomial infections pose a significant global health concern. This study focused on examining the lipid profiles of both non-resistant and clinically resistant strains of Staphylococcus aureus (MRSA 1418), E. coli (ESBL 1384), and Acinetobacter 1379. The main aim was to investigate the relationship between lipid profiles, hydrophobicity, and antibiotic resistance so as to identify the pathogenic potential and resistance factors of strains isolated from patients with sepsis and urinary tract infections (UTIs). The research included various tests, such as antimicrobial susceptibility assays following CLSI guidelines, biochemical tests, biofilm assays, and hydrophobicity assays. Additionally, gas chromatography mass spectrometry (GC-MS) and GC-Flame Ionization Detector (GC-FID) analysis were used for lipid profiling and composition. The clinically isolated resistant strains (MRSA-1418, ESBL-1384, and Acinetobacter 1379) demonstrated resistance phenotypes of 81.80%, 27.6%, and 63.6%, respectively, with a multiple antibiotic resistance index of 0.81, 0.27, and 0.63. Notably, the MRSA-1418 strain, which exhibited resistance, showed significantly higher levels of hemolysin, cell surface hydrophobicity, biofilm index, and a self-aggregative phenotype compared to the non-resistant strains. Gene expression analysis using quantitative real-time PCR (qPCR). Indicated elevated expression levels of intercellular adhesion biofilm-related genes (icaA, icaC, and icaD) in MRSA-1418 (pgaA, pgaC, and pgaB) and Acinetobacter 1379 after 24 h compared to non-resistant strains. Scanning electron microscopy (SEM) was employed for structural investigation. These findings provide valuable insights into the role of biofilms in antibiotic resistance and suggest potential target pathways for combating antibiotic-resistant bacteria.

Modulation of gut-microbiota through probiotics and dietary interventions to improve host health.

Dietary patterns play an important role in regards to the modulation and control of the gut microbiome composition and function. The interaction between diet and microbiota plays an important role in order to maintain intestinal homeostasis, which ultimately affect the host's health. Diet directly impacts the microbes that inhabit the gastrointestinal tract (GIT), which then contributes to the production of secondary metabolites, such as short-chain fatty acids, neurotransmitters, and antimicrobial peptides. Dietary consumption with genetically modified probiotics can be the best vaccine delivery vector and protect cells from various illnesses. A holistic approach to disease prevention, treatment, and management takes these intrinsically linked diet-microbes, microbe-microbe interactions, and microbe-host interactions into account. Dietary components, such as fiber can modulate beneficial gut microbiota, and they have resulting ameliorative effects against metabolic disorders. Medical interventions, such as antibiotic drugs can conversely have detrimental effects on gut microbiota by disputing the balance between Bacteroides and firmicute, which contribute to continuing disease states. We summarize the known effects of various dietary components, such as fibers, carbohydrates, fatty acids, vitamins, minerals, proteins, phenolic acids, and antibiotics on the composition of the gut microbiota in this article in addition to the beneficial effect of genetically modified probiotics and consequentially their role in regards to shaping human health. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Kluyveromyces marxianus MTCC 1389 Augments Multi-stress Tolerance After Adaptation to Ethanol Stress.

During fermentation, yeast cells undergo various stresses that inhibit cell growth and ethanol production. Therefore, the ability to tolerate multiple stresses during fermentation is one of the important characteristics for yeast cells that can be used for commercial ethanol production. In the present study, we evaluated the multi-stress tolerance of parent and ethanol adapted Kluyveromyces marxianus MTCC1389 and their relative gene expression analysis. Multi-stress tolerance was confirmed by determining its cell viability, growth, and spot assay under oxidative, osmotic, thermal, and ethanol stress. During oxidative (0.8% H2O2) and osmotic stress (2 M NaCl), there was significant cell viability of 90% and 50%, respectively, by adapted strain. On the other hand, under 45 °C of thermal stress, the adapted strain was 80% viable while the parent strain was 60%. In gene expression analysis, the ethanol stress responsive gene ETP1 was significantly upregulated by 3.5 folds, the osmotic stress gene SLN1 was expressed by 3 folds, and the thermal stress responsive gene MSN2 was expressed by 7 folds. This study shows adaptive evolution for ethanol stress can develop other stress tolerances by changing relative gene expression of osmotic, oxidative, and thermal stress responsive genes.

Draft genome sequence of methicillin-resistant Staphylococcus aureus strain D1418m22 isolated from human wound pus.

This study reveals the complete genome sequence of methicillin-resistant Staphylococcus aureus (MRSA) strain d1418m22, sourced from a Karnal, Haryana, human skin wound. Classified as community-associated MRSA, it features a 2.78-MB genome harboring Staphylococcus-specific genetic elements, encompassing 2,625 protein-coding genes and 18 antimicrobial resistance genes.

"In vitro antimicrobial and synergistic effect of fermented Indian zebu (Sahiwal) cow colostrum whey derived peptides with Lactobacillus rhamnosus against pathogenic bacteria".

Bioactive peptides (BAPs) have been found to promote health through various mechanisms. Among them, antimicrobial peptides are gaining recognition as promising novel treatments. This study aims to generate BAPs from bovine colostrum whey using the proteolytic activity of Lactobacillus rhamnosus C25 and to evaluate their potential antibacterial efficacy, including their ability to synergistic efficacy against resistant bacteria. Bioactive peptides were successfully generated from lactobacillus culture proteases that were cultivated through batch fermentation. The resulting peptide fractions were then evaluated for their antibacterial efficacy against a selection of strains, including E. coli ATCC25922, S. aureus MTCC1144, Acinetobacter baumannii ATCC 17978, as well as clinically isolated resistant strains of E. coli (ESBL 1384), Acinetobacter 1379, and S. aureus (MRSA 1418). Notably, the peptide fractions with a molecular weight of < 10 kDa (0-10 kDa) significantly increased the membrane permeability of both E. coli (70.30 ± 0.41%) and S. aureus (63.04 ± 0.31%) as assessed by the crystal violet assay. The checkerboard method was utilized to perform synergistic tests with peptides and antibiotics. The peptide fractions with a molecular weight of (< 10 kDa) demonstrated synergistic effects with several antibiotics, including gentamycin, Rifampicin, Levofloxacin, Ciprofloxacin, and Chloramphenicol, against the resistant ESBL 1384 strain, as indicated by ΣFICI values of 0.55, 0.53, 0.52, 0.54, and 0.52, respectively. Furthermore, the HT-29 cell line remained completely unaffected by both peptide fractions. These findings suggest that the < 10 kDa peptide fraction possesses significant antibacterial efficacy against both reference and ESBL 1384 resistant bacterial strain. Additionally, both MRSA 1418 and Acinetobacter 1379 displayed resistance to all fractions tested. To summarize the findings of this study, colostrum whey peptides with a broad spectrum of antimicrobial activity can be efficiently produced through fermentation. This method could prove valuable for both the pharmaceutical and food industries. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-023-05776-2.

Evaluation of the antimicrobial attribute of bioactive peptides derived from colostrum whey fermented by Lactobacillus against diarrheagenic E. coli strains.

Colostrum known as "liquid gold" contains approximately 60-80% of whey proteins that can be a great source of bioactive peptide production. Therefore, this study aimed to perform a comparative antimicrobial evaluation of the bioactive peptide generated from L. rhamnosus C25, L. rhamnosus C6, and L. casei NCDC17 fermented colostrum whey. Peptide fractions 10 kDa, 5 kDa, and 3 kDa were isolated using their respective molecular weight cut-off membranes and antimicrobial activity was evaluated against diarrheagenic E. coli strains. The higher inhibition was shown by < 10 kDa peptide fractions from L. rhamnosus C25 fermented colostrum whey and the zone of inhibition was 15 ± 0.06 (E. coli MTCC 723), 17 ± 0.04 (E. coli MTCC 724), 18 ± 0.05 (E. coli MTCC 725), and 17 ± 0.02 (E. coli ATCC 25922). In addition, ST-1 and LT-1 genes of E. coli strains were also confirmed using PCR which is responsible for the diarrheagenic property. Further, the interaction of potent peptides against E. coli strains was also observed by scanning electron microscope. Hence, the significance of the present study emphasized that these bioactive peptides generated from fermented colostrum whey can be used as ingredients in functional food against diarrhoea.

Peptidomics-based identification of an antimicrobial peptide derived from goat milk fermented by Lactobacillus rhamnosus (C25).

Antimicrobial peptides (AMPs) are emerging as promising novel drug applicants. In the present study, goat milk was fermented using Lactobacillus rhamnosus C25 to generate bioactive peptides (BAPs). The peptide fractions generated were separated using ultrafiltration membranes with molecular weight cut-offs of 3, 5, and 10 kDa, and their antimicrobial activity toward Gram-positive and Gram-negative bacteria was investigated. Isolated AMPs were characterized using RP-HPLC and identified by LC-MS/MS. A total of 569 sequences of peptides were identified by mass spectrometry. Out of the 569, 36 were predicted as AMPs, 21 were predicted as cationic, and out of 21, 6 AMPs were helical peptides. In silico analysis indicated that the majority of peptides were antimicrobial and cationic in nature, an important factor for peptide interaction with the negative charge membrane of bacteria. The results showed that the peptides of <5 kDa exhibited maximum antibacterial activity against E. faecalis, E. coli, and S. typhi. Further, molecular docking was used to evaluate the potent MurD ligase inhibitors. On the basis of ligand binding energy, six predicted AMPs were selected and then analyzed by AutoDock tools. Among the six AMPs, peptides IGHFKLIFSLLRV (-7.5 kcal/mol) and KSFCPAPVAPPPPT (-7.6 kcal/mol), were predicted as a high-potent antimicrobial. Based on these findings, in silico investigations reveal that proteins of goat milk are a potential source of AMPs. This is for the first time that the antimicrobial peptides produced by Lactobacillus rhamnosus (C25) fermentation of goat milk have been identified via LC-MS/MS and predicted as AMPs, cationic charges, helical structure in nature, and potent MurD ligase inhibitors. These peptides can be synthesized and improved for use as antimicrobial agents. PRACTICAL APPLICATIONS: Goat milk is considered a high-quality source of milk protein. According to this study, goat milk protein is a potential source of AMPs, Fermentation can yield goat milk-derived peptides with a broad antibacterial activity spectrum at a low cost. The approach described here could be beneficial in that the significant AMPs can be synthesized and used in the pharmaceutical and food industries.

Antimicrobial bioactive peptides from goat Milk proteins: In silico prediction and analysis.

The main goal of this study was to assess the potential proteins of goat milk (i.e. α-s1-casein, α-s2-casein, ß-casein, κ-casein, α-lactoglobulin and ß-lactalbumin) as precursors of antimicrobial peptides (AMPs). Bioinformatics tools such as BIOPEP-UWM (enzyme action) were used for the in silico gastrointestinal digestion via a cocktail of pepsin, trypsin, and chymotrypsin A. The antimicrobial activity of peptides was predicted by using four algorithms, including Random Forest, Support Vector Machines, Artificial Neural Network and Discriminant Analysis on CAMPR3 online server, which works on Hidden Markov Models. Different online tools predicted the physiochemical properties, allergenicity, and toxicity of peptides as well. In silico gastrointestinal digestion simulation of proteins by enzymes cocktail yielded a total of 83 potential AMPs, with thirteen peptides being confident by all four algorithms. More AMPs were released from ß-casein (21) than from ß-lactoglobulin (16), α-s1-casein (15), α-s2-casein (12), κ-casein (11) and α-lactalbumin (9). A total of 17 peptides were cationic, and the majority of the peptides were extended AMPs. These peptides were released from α-s1-casein (SGK, IQK), α-s2-casein (SIR, AIH, TQPK), ß-casein (GPVR, AVPQR, AIAR, GVPK, SQPK, PVPQK, IH, VPK), k-casein (AIPPK, QQR, IAK, TVPAK). All of the AMPs were anticipated to be non-toxic, and 54 of the 83 peptides were confirmed to be non-allergic, with the remaining 29 suspected of being allergenic and 31 to be predicted to have good water solubility. Further the molecular docking was used to evaluate the potent dihydropteroate synthase (DHPS) inhibitors. On the basis of ligand binding energy, 17 predicted AMPs were selected and then analyzed by AutoDock tools. Among the 17 AMPs, 3 AMPs were predicted as high-potent antimicrobial. Based on these findings, in silico investigations reveal that proteins of goat milk are a potential source of AMPs. These peptides can be synthesized and improved for use in the food sector. PRACTICAL APPLICATIONS: Goat milk is regarded as a high-quality milk protein source. According to this study, goat milk protein is a possible source of AMPs, and therefore, most important AMPs can be synthesized and developed for use in the food sector.

In silico identification of antidiabetic and hypotensive potential bioactive peptides from the sheep milk proteins-a molecular docking study.

An in silico approach was used for hydrolysis of sheep milk proteins (α-s1, α-s2, ß-casein, κ-Cn, α-lactalbumin, and ß-lactoglobulin) by gastrointestinal enzymes in order to generate bioactive peptides (BAPs) that can inhibit ACE and DPP-IV. Sheep milk proteins showed higher similarity with goat milk proteins. These data were acquired via the Clustal Omega tool to perform sequence alignment analysis. The BIOPEP-UWM database was used to examine the ability of sheep milk protein sequences to generate BAPs, which included a description of their potential bioactivity as well as the frequency of fragments with specified activities. Using the "Enzyme(s) action" tool (BIOPEP-UWM), digestive enzymes pepsin, trypsin, and chymotrypsin, and three enzyme combinations were selected to computationally hydrolyze milk proteins for obtaining information about ACE and DPP-IV inhibitory peptides. Other online programs were used to test potential peptides for bioactivity, toxicity, and physicochemical properties. BAPs produced from PTC-hydrolyzed proteins were analyzed using a peptide ranker, and their inhibitory effects on ACE and DPP-IV were determined using molecular docking. Consequently, the results of molecular docking analysis show that the peptide PSGAW (αS1-Cn f155-159) binds to DPP-IV with binding energy (-8.9 kcal/mol). But in the case of ACE, two potential BAPs were selected: QPPQPL (ß-Cn f161-166) and PSGAW. These two BAPs revealed a higher binding affinity for ACE with a binding energy of -9.8 kcal/mol. Thus, the results showed that sheep milk proteins were a promising source of antidiabetic and hypotensive peptides. However, experimental and pre-clinical studies are necessary to assay their therapeutic effects. PRACTICAL APPLICATIONS: Sheep milk proteins are known as a high-quality milk protein resource. Effective enzymatic hydrolysis of sheep milk proteins can release bioactive peptides and also release potential ACE and DPP-IV inhibitory peptides. This in silico study specifies a theoretical root for sheep milk proteins as a novel source of potential bioactive peptides and may offer guidance for invitro hydrolysis of proteins for the production of bioactive peptides valuable for human consumption.

Etiology and management of Alzheimer's disease: Potential role of gut microbiota modulation with probiotics supplementation.

Alzheimer's disease (AD) is the leading type of dementia in aging people and is a progressive condition that causes neurodegeneration, resulting in confusion, memory loss, and deterioration of mental functions. AD happens because of abnormal twisting of the microtubule tau protein in neurons into a tangled neurofibrillary structure. Different factors responsible for AD pathogenesis include heavy metals, aging, cardiovascular disease, and environmental and genetic factors. Market available drugs for AD have several side effects that include hepato-toxicity, accelerated cognitive decline, worsened neuropsychiatric symptoms, and triggered suicidal ideation. Therefore, an emerging alternative therapeutic approach is probiotics, which can improve AD by modulating the gut-brain axis. Probiotics modulate different neurochemical pathways by regulating the signalling pathways associated with inflammation, histone deacetylation, and microglial cell activation and maturation. In addition, probiotics-derived metabolites (i.e., short-chain fatty acid, neurotransmitters, and antioxidants) have shown ameliorative effects against AD. Probiotics also modulate gut microbiota, with a beneficial impact on neural signalling and cognitive activity, which can attenuate AD progression. Therefore, the current review describes the etiology and mechanism of AD progression as well as various treatment options with a focus on the use of probiotics. PRACTICAL APPLICATIONS: In an aging population, dementia concerns are quite prevalent globally. AD is one of the most commonly occurring cognition disorders, which is linked to diminished brain functions. Scientific evidence supports the findings that probiotics and gut microbiota can regulate/modulate brain functions, one of the finest strategies to alleviate such disorders through the gut-brain axis. Thus, gut microbiota modulation, especially through probiotic supplementation, could become an effective solution to ameliorate AD.

Physiological growth and galactose utilization by dairy yeast Kluyveromyces marxianus in mixed sugars and whey during fermentation.

The dairy yeast Kluyveromyces marxianus represents a promising industrial strain useful for the production of bioethanol from cheese whey. Physiology of the five K. marxianus strains on galactose was examined during batch cultivation under controlled aerobic conditions on minimal media and one of the strains designated K. marxianus strain 6C17 which presented the highest specific galactose consumption rate. A maximum specific growth rate of 0.34 and 0.37 h-1, respectively, was achieved using batch cultivation in a minimal medium and a complex medium amended with galactose (50 g/L) at 37 °C. The sugar was metabolized for the production of ethanol as the chief metabolite with a maximum ethanol yield of 0.39 g/g of galactose. Different growth behaviors were observed when galactose was used with other sugar such as glucose, lactose and fructose. The growth rates on hydrolyzed cheese whey were also measured, and a maximum specific growth rate of 0.39 and 0.32 h-1 was observed with glucose and galactose, respectively, with the maximum flux diverted toward ethanol production. This approach of studying the physiology of thermotolerant K. marxianus on hydrolyzed whey during fermentation would be helpful in achieving higher yields of ethanol.

Comparative Analysis of Oxidative Stress During Aging of Kluyveromyces marxianus in Synthetic and Whey Media.

During the aging of yeast culture, Kluyveromyces marxianus undergoes a number of changes in physiology and these changes play a significant role during fermentation. Aged stationary phase cells were found to contain more reactive oxygen species. Additionally, the level of oxidant is counteracted by the antioxidant defense system of the cells. Comparison of 3-day-old culture of K. marxianus with 45-day stationary phase culture represents an increased level of ROS inside the cells. Moreover, a decrease in glutathione content was observed over the set of the incubation period. The increased level of superoxide dismutase (SOD) and catalase also revealed that there is oxidative stress during the long period incubation of the stationary phase cells of K. marxianus. The actual phenomenon of aging in dairy yeast K. marxianus is a complex process, but the present study signifies that role of antioxidant defense system during aging in stationary phase cells of K. marxianus.

Utilization of Cheese Whey Using Synergistic Immobilization of ß-Galactosidase and Saccharomyces cerevisiae Cells in Dual Matrices.

Whey is a byproduct of the dairy industry, which has prospects of using as a source for production of various valuable compounds. The lactose present in whey is considered as an environmental pollutant and its utilization for enzyme and fuel production, may be effective for whey bioremediation. The dairy yeast Kluyveromyces marxianus have the ability to utilize lactose sharply as the major carbon source for the production of the enzyme. Five strains were tested for the production of the ß-galactosidase using whey. The maximum ß-galactosidase activity of 1.74 IU/mg dry weight was achieved in whey using K. marxianus MTCC 1389. The biocatalyst was further immobilized on chitosan macroparticles and exhibited excellent functional activity at 35 °C. Almost 89 % lactose hydrolysis was attained for concentrated whey (100 g/L) and retained 89 % catalytic activity after 15 cycles of reuse. Finally, ß-galactosidase was immobilized on chitosan and Saccharomyces cerevisiae on calcium alginate, and both were used together for the production of ethanol from concentrated whey. Maximal ethanol titer of 28.9 g/L was achieved during fermentation at 35 °C. The conclusions generated by employing two different matrices will be beneficial for the future modeling using engineered S. cerevisiae in scale-up studies.

ß-Glucosidase activity and bioconversion of isoflavones during fermentation of soymilk.

BACKGROUND: Lactobacillus rhamnosus C6 strain showed higher ß-glucosidase activity as well as biotransformation of isoflavones from glycones (daidzin and genistin) to aglycones (daidzein and genistein) in soymilk. However, L. rhamnosus C2 and Lactobacillus casei NCDC297 also exhibited similar activity during soymilk fermentation. These three strains can be selected for the development of functional fermented soy foods enriched with aglycone forms of isoflavones, such as soy yoghurt, soy cheese, soy beverages and soy dahi. RESULTS: The study determined ß-glucosidase activity of probiotic Lactobacillus cultures for bioconversion of isoflavones to aglycones in fermenting soymilk medium. Soymilk was fermented with six strains (L. rhamnosus C6 and C2, L. rhamnosus NCDC19 and NCDC24 and L. casei NCDC17 and NCDC297) at 37 °C for 12 h. The highest ß-glucosidase activity and isoflavone bioconversion after 12 h occurred by L. rhamnosus C6 culture during fermentation in soymilk. Increased isoflavone aglycone content in fermented soymilk is likely to improve the biological functionality of soymilk (e.g. antioxidant activity, alleviation of hormonal disorders in postmenopausal women, etc.). CONCLUSION: Lactobacillus rhamnosus C6 culture can be used for the development of functional fermented soy-based products.

Functional significance of bioactive peptides derived from soybean.

Biologically active peptides play an important role in metabolic regulation and modulation. Several studies have shown that during gastrointestinal digestion, food processing and microbial proteolysis of various animals and plant proteins, small peptides can be released which possess biofunctional properties. These peptides are to prove potential health-enhancing nutraceutical for food and pharmaceutical applications. The beneficial health effects of bioactive peptides may be several like antihypertensive, antioxidative, antiobesity, immunomodulatory, antidiabetic, hypocholesterolemic and anticancer. Soybeans, one of the most abundant plant sources of dietary protein, contain 36-56% of protein. Recent studies showed that soy milk, an aqueous extract of soybean, and its fermented product have great biological properties and are a good source of bioactive peptides. This review focuses on bioactive peptides derived from soybean; we illustrate their production and biofunctional attributes.

Nisin and class IIa bacteriocin resistance among Listeria and other foodborne pathogens and spoilage bacteria.

Food safety has been an important issue globally due to increasing foodborne diseases and change in food habits. To inactivate foodborne pathogens, various novel technologies such as biopreservation systems have been studied. Bacteriocins are ribosomally synthesized peptides or proteins with antimicrobial activity produced by different groups of bacteria, but the bacteriocins produced by many lactic acid bacteria offer potential applications in food preservation. The use of bacteriocins in the food industry can help reduce the addition of chemical preservatives as well as the intensity of heat treatments, resulting in foods that are more naturally preserved. However, the development of highly tolerant and/or resistant strains may decrease the efficiency of bacteriocins as biopreservatives. Several mechanisms of bacteriocin resistance development have been proposed among various foodborne pathogens. The acquiring of resistance to bacteriocins can significantly affect physiological activity profile of bacteria, alter cell-envelope lipid composition, and also modify the antibiotic susceptibility/resistance profile of bacteria. This article presents a brief review on the scientific research about the various possible mechanisms involved in the development of resistance to nisin and Class IIa bacteriocins among the foodborne pathogens.

Safety assessment and evaluation of probiotic potential of bacteriocinogenic Enterococcus faecium KH 24 strain under in vitro and in vivo conditions.

In the present investigation, a previously isolated Enterococcus faecium KH 24 strain was evaluated for the presence of virulence determinants (agg, esp, efaAfm, gelE, cylA, cylB, clyM, cpd, cob, ccf, ace and hyl), sensitivity to various antibiotics and production of biogenic amines. No virulence determinants were detected, except efaAfm. KH 24 was found to be sensitive to most of the tested antibiotics and none of the biogenic amines were produced by it. Moreover, KH 24 showed good in vitro tolerance to biological barriers and furthermore, its survival in gut of mice was also evaluated. Mice group fed with E. faecium KH 24 strain showed better weight gain and nearly 1 log cfu/g decrease in Salmonella enteritidis counts in the intestines as compared to control (p<0.05). Enhanced growth of lactobacilli (p<0.05) and decrease in coliform counts (p<0.05) were also observed in test group. E. faecium KH 24 is, therefore, found to be a safe strain and it may be used as protective culture or as a probiotic in food preparations.

Differential effects of hypoxia-ischemia on phosphorylation of the N-methyl-D-aspartate receptor in one- and three-week-old rats.

The effects of transient cerebral hypoxia-ischemia (HI) on phosphorylation of the NR1 subunit of the N-methyl-D-aspartate (NMDA) receptor were investigated in 7 (P7)- and 21 (P21)-day-old rats. Unilateral HI was induced by ligation of the right common carotid artery and exposure to 8% O(2)/92% N(2) for 120 (P7) or 90 (P21) min. Phosphorylation by protein kinase A (PKA; S897) and PKC (S896 and S890) was depressed in the ipsilateral hemisphere relative to both naïve controls and the contralateral hemisphere immediately following HI at both ages. At P7, but not P21, reperfusion resulted in an initial recovery to control phosphorylation levels at all 3 sites followed by a secondary decline. At both ages, pS896 was less than control values after 24 h of recovery, whereas pS890 had returned to control levels by this time. pS897 recovered to control levels by 24 h in P21 animals but not in P7 animals. Differential effects of HI on phosphorylation of the NMDA receptor at P7 and P21 may contribute to age-related changes in sensitivity to HI.

Trapping-induced changes in expression of the N-methyl-D-aspartate receptor in the hippocampus of snowshoe hares.

Live-trapping of animals in natural populations is one of the main ways to determine population processes. We examined the effects of live-trapping on the expression of N-methyl-aspartate (NMDA) receptor subunits in the hippocampus of snowshoe hares. Snowshoe hares were obtained either with or without the stress of live-trapping. The CA1, CA3 and dentate gyrus were dissected and analyzed for the presence of NMDA receptor subunits. Trapping resulted in a significant reduction of NMDA receptor 1 (NR1) in each of the regions examined but did not affect the levels of either NMDA receptor 2A or B (NR2A or NR2B). Co-immunoprecipitation analysis showed that the association between NR1 and NR2A was decreased in the trapped animals. These results suggest that stress associated with the trapping experience may adversely affect the structure and/or function of the NMDA receptor.