Amylase Research Articles

Purification of ursolic acid and β-sitosterol from endophytic Alternaria alternata for their alpha-amylase inhibitory activity

Fungal endophytes are a known warehouse of bioactive compounds with multifarious applications. In the present investigation two compounds, β-Sitosterol (1) and ursolic acid (2), were isolated from Alternaria alternata, an endophytic fungus associated with Morus alba Linn for the first time. The structure of the compounds was elucidated on the basis of comprehensive spectral analysis (UV, IR, 1 H-, 13 C- and 2D-NMR, as well as HRESI–MS). In the in vitro alpha amylase inhibitory assay both compounds (1) and (2) show potent antidiabetic activity. In support, Docking studies indicate significant binding affinity of the isolated compounds. Hence from the present study, it can be concluded that endophytic fungi in Morus alba Linn can find use in antidiabetic drug development in the medicinal industry. Communicated by Ramaswamy H. Sarma

The effects of eugenol on histological, enzymatic, and oxidative parameters in the major salivary glands and pancreas of healthy male Wistar rats

ObjectiveWe evaluated the effects of eugenol on histological, enzymatic, and oxidative parameters in the pancreas, parotid, submandibular, and sublingual glands of healthy male rats. DesignTwenty-four adult Wistar rats were assigned into four groups (n = 6/group). Control rats received 2% Tween-20 (eugenol vehicle), whereas the other animals received 10, 20, and 40 mg kg−1 eugenol through gavage daily for 60 d. Major salivary and pancreatic glands were weighed and preserved fixed for microscopic analysis and frozen for in vitro assays. ResultsEugenol did not alter glands’ weight and serum amylase activity regardless of the concentration. The highest dose of eugenol caused an increase in pancreatic amylase activity and a reduction of lipase activity from serum and pancreas. Eugenol at 40 mg kg−1 diminished the activity of SOD and FRAP in the submandibular gland and CAT and FRAP in the sublingual gland. However, it did not exert any effect on GST regardless of the gland. Additionally, 40 mg kg−1 eugenol increased MDA levels in pancreatic, parotid, and submandibular glands and NO levels in the sublingual. The concentrations of eugenol induced distinct responses in the glands regarding the activity of Na+/K+, Mg2+, and total ATPase activity. They also affected histomorphometrical and histochemistrical parameters in the submandibular gland only. ConclusionsResults indicated that 40 mg kg−1 eugenol altered most of the biochemical and oxidatived parameters of digestive glands. Only submandibular glands presented histological changes after eugenol exposure suggesting potential implications for its function.

COMPARATIVE ANALYSIS OF THE GENETIC STRUCTURE OF SCALED AND FRAMED CARP CHERNIGIVRYBHOSP PJSC

Aim. This study aimed to analyze the genetic structure of Ukrainian scaled and framed carp, by examining the frequencies of allelic variants and genotypes of protein polymorphic systems. Methods of research. The research was conducted using electrophoresis in polyacrylamide and starch gels, followed by histochemical staining. The genetic structure of the carp groups was assessed by analyzing the distribution of allele and genotype frequencies of loci encoding various proteins and enzymes: transferrin (TF), albumin (ALB), esterase (EST), hemoglobin (HB), ceruloplasmin (С), amylase (AM) and purine nucleoside phosphorylase (PN). The obtained data were mathematically processed using the "BIOSYS" software. Research results. In the scaled and framed studied carp groups, the spectra of hemoglobin, ceruloplasmin, amylase and purine nucleoside phosphorylase, which encode the corresponding enzymes were found to be monomorphic under the given conditions of electrophoretic investigation. This finding was confirmed by the data of other authors. The genetic structure of scaled and framed carp. was analyzed using the transferrin, albumin, and esterase loci. Four allelic forms of the transferrin locus were found in the investigated stock of scaled carp: Tf A, Tf B, Tf C1 and Tf C2. The highest frequency was observed for the Tf B allelic variant (0.400), while the lowest frequency was observed for Tf A (0.050). Four allelic forms of the transferrin locus have been established in the framed carp group: Tf B, Tf C1, Tf C2 and Tf D. The allelic variant B occurred with the highest frequency (0.739), while the Tf C2 allele occurred with the lowest frequency (0.043). The analysis of the genotypes of the stock of scaled carp showed that only 5 of the 15 possible genotypes were present, among which the homozygous genotype BB occurred with the highest frequency, and it was 40%. The genotypes AA, AB, AC2, AD, BC1, BC2, BD, C1D, C2D and DD were absent. In this group of framed carp, three genotypes out of the fifteen possible variations were found namely Tf BВ, Tf С1С2 and Tf С1D. The homozygous BB genotype occurred with the highest frequency, accounting for 73.9%. According to the esterase locus, a specific feature of both scaled and framed carp studied stocks on this farm was the absence of the SS homozygous genotype. In the studied scaled carp group, the FS heterozygous genotype had the highest frequency (65%). The Est F allele had a frequency of 0.675, while the Est S allele had a frequency of 0.261. In the electrophoretic analysis of blood plasma in framed carp, the Est F allelic variant occurred with a frequency of 0.739, while the Est S allele occurred with a frequency of 0.261. The FS heterozygous genotype in the framed carp stock had a predominant frequency of 52.2%. At the albumin locus in both scaled and framed carp groups, two alleles, A and B, were identified, with the frequencies 0.600 and 0.400 in scaled and 0.848 and 0.152 in framed carp, respectively. Three possible genotypes of the albumin locus were identified. According to the analysis of the correspondence of the distribution of genotypes of scaled and framed carp to the ratio according to the Hardy-Weinberg law at a significance level of 5%, it was shown that there are no deviations at the EST and ALB loci. According to the locus of transferrin in both studied scaled and framed carp stocks, the presence of a statistically significant difference in the observed distribution of the number of genotypes compared to the expected according to the Hardy-Weinberg law was established. In the scaled carp group, the observed level of heterozygosity at the transferrin locus was lower (0.300) than expected (0.692). The level of observed heterozygosity was observed for the esterase and albumin loci, which exceeded the expected values in both cases. In the framed carp of the studied group, the highest level of observed heterozygosity was found at the EST locus (0.522), while the lowest was observed at the TF locus (0.261). According to the ALB locus, the level of observed heterozygosity was 0.304. In contrast to the transferrin locus, the observed heterozygosity values for the esterase and albumin loci in this group exceeded the expected values. The average values of heterozygosity for the three loci were higher in the scaled carp group (Ho = 0.517 and He = 0.545) compared to the framed group (Ho = 0.362 and He = 0.365). Conclusions. The studied protein and enzyme systems allowed for the demonstration of the genetic structure features of scaled and framed carp farmed by Chernihivrybhosp PJSC, which were expressed in a specific set of TF allelic variants. Another peculiarity of individuals from this farm was the absence of the SS genotype at the esterase locus.

Potencial of Aloin B Compound and its Derivatives as Type-2 Antidiabetic

Type-2 diabetes mellitus occurs due to suboptimal insulin function (insulin resistance) or decreased insulin function. Type-2 diabetes mellitus treatment is chronic and lifelong. One of the treatments is the use of insulin and oral anti-diabetic drugs. This treatment requires a long period of time and can cause unwanted side effects. Therefore, alternative treatments are needed with minimal side effects by utilizing herbal plants containing Aloin B compounds because they have been proven to be used as antidiabetic agents. These compounds can be found from the Aloe Vera plant (Aloe Vera l.). The aim of this study was to find compounds derived from Aloin B compounds that have the most potential as anti-diabetic type-2 by inhibiting the pancreatic α-amylase enzyme (code: 1B2Y) in breaking down starch in the body. The certainty of the presence of the compound Aloin B in the flesh of the aloe plant was confirmed by the LC-MS test. This research was conducted using the Quantitative Structure-Activity Relationship (QSAR) and Molecular Bonding method. The results showed that the ID S22 compound with the IUPAC name (S)-10-amino-1,2,8-trihydroxy-6- (hydroxymethyl) -10- ((2R,3R,4S,5S,6R) -2,3, 4,5-tetrahydroxy–6(hydroxymethyl) tetrahydro-2H-piran-2-yl) anthracene-9(10H)-one is the most potent compound from Aloin B derivatives as a type-2 antidiabetic agent in the mechanism of inhibiting α-enzyme action pancreatic amylase, based on the value of R2 = 0.980, the PRESS value of the compound was 0.0004, the binding energy value was -7.07 kcal/mol, the inhibition constant was 6.58 uM and the formation of hydrogen bonds between the compound and the amino acid residues aspirin, glycine, threonine and arginine.

LC-MS/MS investigation of phytochemical ingredients and alpha amylase inhibition activity of Cassia siamea Lam leaves aqueous extracts

LC-MS/MS investigation of phytochemical ingredients and alpha amylase inhibition activity of Cassia siamea Lam leaves aqueous extracts

Green Synthesis and Characterization of Novel Silver Nanoparticles Using Achillea maritima subsp. maritima Aqueous Extract: Antioxidant and Antidiabetic Potential and Effect on Virulence Mechanisms of Bacterial and Fungal Pathogens

Novel silver nanoparticles were synthesized based on a simple and non-toxic method by applying the green synthesis technique, using, for the first time, the aqueous extract of an extremophile plant belonging to the Achillea maritima subsp. maritima species. AgNP characterization was performed via UV-Visible, front-face fluorescence spectroscopy, and FTIR and XRD analyses. AgNP formation was immediately confirmed by a color change from yellow to brown and by a surface plasmon resonance peak using UV-Vis spectroscopy at 420 nm. The biosynthesized AgNPs were spherical in shape with a size ranging from approximatively 14.13 to 21.26 nm. The presented silver nanoparticles exhibited strong antioxidant activity following a DPPH assay compared to ascorbic acid, with IC50 values of about 0.089 µg/mL and 22.54 µg/mL, respectively. The AgNPs showed higher antidiabetic capacities than acarbose, by inhibiting both alpha amylase and alpha glucosidase. The silver nanoparticles could affect various bacterial mechanisms of virulence, such as EPS production, biofilm formation and DNA damage. The silver nanoparticles showed no lysozyme activity on the cell walls of Gram-positive bacteria. The AgNPs also had a strong inhibitory effect on the Candida albicans virulence factor (extracellular enzymes, biofilm formation). The microscopic observation showed abnormal morphogenesis and agglomeration of Candida albicans exposed to AgNPs. The AgNPs showed no cytotoxic effect on human cells in an MTT assay. The use of novel silver nanoparticles is encouraged in the formulation of natural antimicrobial and antidiabetic agents.

Biogenic Fabrication of Iron Oxide Nanoparticles from Leptolyngbya sp. L-2 and Multiple In Vitro Pharmacogenetic Properties.

Metallic nanoparticles have received a significant amount of reflection over a period of time, attributed to their electronic, specific surface area, and surface atom properties. The biogenic synthesis of iron oxide nanoparticles (FeONPs) is demonstrated in this study. The green synthesis of metallic nanoparticles (NPs) is acquiring considerable attention due to its environmental and economic superiorities over other methods. Leptolyngbya sp. L-2 extract was employed as a reducing agent, and iron chloride hexahydrate (FeCl3·6H2O) was used as a substrate for the biogenic synthesis of FeONPs. Different spectral methods were used for the characterization of the biosynthesized FeONPs, ultraviolet-visible (UV-Vis) spectroscopy gave a surface plasmon resonance (SPR) peak of FeONPs at 300 nm; Fourier transform infrared (FTIR) spectral analysis was conducted to identify the functional groups responsible for both the stability and synthesis of FeONPs. The morphology of the FeONPs was investigated using scanning electron microscopy (SEM), which shows a nearly spherical shape, and an X-ray diffraction (XRD) study demonstrated their crystalline nature with a calculated crystallinity size of 23 nm. The zeta potential (ZP) and dynamic light scattering (DLS) measurements of FeONPs revealed values of -8.50 mV, suggesting appropriate physical stability. Comprehensive in-vitro pharmacogenetic properties revealed that FeONPs have significant therapeutic potential. FeONPs have been reported to have potential antibacterial and antifungal properties. Dose-dependent cytotoxic activity was shown against Leishmania tropica promastigotes (IC50: 10.73 µg/mL) and amastigotes (IC50: 16.98 µg/mL) using various concentrations of FeONPs. The cytotoxic potential was also investigated using brine shrimps, and their IC50 value was determined to be 34.19 µg/mL. FeONPs showed significant antioxidant results (DPPH: 54.7%, TRP: 49.2%, TAC: 44.5%), protein kinase (IC50: 96.23 µg/mL), and alpha amylase (IC50: 3745 µg/mL). The biosafety of FeONPs was validated by biocompatibility tests using macrophages (IC50: 918.1 µg/mL) and red blood cells (IC50: 2921 µg/mL). In conclusion, biogenic FeONPs have shown potential biomedical properties and should be the focus of more studies to increase their nano-pharmacological significance for biological applications.

A COMPUTATIONAL GENOME ANALYSIS OF STRAIN BACILLUS SUBTILIS MIZ-8 ISOLATED FROM BEKANG REVEALS A DISTINCT CHROMOSOME AND PLASMID CONFERRING SELECTIVE ADVANTAGE

Bacillus subtilis Miz-8 is isolated from Bekang, a traditional fermented soybean food of Mizoram, India. Here, we report computationally analyzed strain Miz-8 mapped to the annotated genome of the reference strain Bacillus subtilis subsp. natto BEST195. Miz-8 has a circular chromosome and a small plasmid with genome size of 4105264 bp and 5838 bp, respectively. Genome contains the genes responsible for synthesis of biosynthetic metabolites like surfactin, fengycin, bacillibactin, bacilysin, subtilosin A and alpha amylase production. The strain Miz-8 harbored virulence genes identical to the strain BEST195 render themselves harmless for human consumption. However, Strain Miz-8 has one intact prophage region but no integrase protein, making it incapable of lateral transfer of antimicrobials. Antibiotic resistance genes were predicted among which tmrB gene was on perfect hit. Plasmid of strain Miz-8 contains no prophage sequences and antibiotic resistance genes. Furthermore, there were several single nucleotide polymorphisms and 344 insertion-deletion polymorphisms. Bacillus subtilis Miz-8's genomic information unmasks its functional significance on human health.

The impact of physical exercise on the consolidation of fear extinction memories.

Based on the mechanisms of fear extinction, exposure therapy is the most common treatment for anxiety disorders. However, extinguished fear responses can reemerge even after successful treatment. Novel interventions enhancing exposure therapy efficacy are therefore critically needed. Physical exercise improves learning and memory and was also shown to enhance extinction processes. This study tested whether physical exercise following fear extinction training improves the consolidation of extinction memories. Sixty healthy men underwent a differential fearconditioning paradigm with fear acquisition training on day 1 and fear extinction training followed by an exercise or resting control intervention on day 2. On day 3, retrieval and reinstatement were tested including two additional but perceptually similar stimuli to explore the generalization of exercise effects. Exercise significantly increased heart rate, salivary alpha amylase, and cortisol, indicating successful exercise manipulation. Contrary to our expectations, exercise did not enhance but rather impaired extinction memory retrieval on the next day, evidenced by significantly stronger differential skin conductance responses (SCRs) and pupil dilation (PD). Importantly, although conditioned fear responses were successfully acquired, they did not fully extinguish, explaining why exercise might have boosted the consolidation of the original fear memory trace instead. Additionally, stronger differential SCRs and PD toward the novel stimuli suggest that the memory enhancing effects of exercise also generalized to perceptually similar stimuli. Together, these findings indicate that physical exercise can facilitate both the long-term retrievability and generalization of extinction memories, but presumably only when extinction was successful in the first place.

Modulatory effect of green coffee bioactives on high-fat diet–induced obesity in C57BL6 mice model

ObjectivesThe aim of the present study was to determine the efficacy of green coffee bioactives in ameliorating the effects of high-fat diet (HFD)-induced obesity through in vitro and in vivo assessments. MethodsGreen coffee extract (GCE) was obtained by implementing a novel green extraction technique. The efficacy of GCE to inhibit in vitro pancreatic amylase and lipase was evaluated. Further, in vivo studies were conducted using a C57BL6 mice model grouped as starch-fed diet control, HFD control, HFD + positive control, HFD + GCE (100 mg/kg body weight), and HFD + GCE (200 mg/kg body weight). Animal body weight, diet intake, and fecal fat excretion were measured during the feeding period. On completion of the experiment, blood serum was collected for biochemical analysis, and organs were harvested for assessing the obesity-related biomarkers. ResultsThe obtained GCE was enriched with polyphenols and alkaloids. GCE led to significant (P < 0.05) in vitro inhibition of pancreatic amylase and lipase. GCE supplementation considerably prevented weight gain in treated groups post-consumption of HFD. It also led to increased fecal fat excretion and regulated the high-fat–mediated blood glucose levels. GCE effectively modulated the blood lipid profile, morphology of adipose and liver tissues, and liver antioxidant defense enzymes and resulted in hepatoprotective effects. It also downregulated the genes associated with lipid biosynthesis. ConclusionsGCE exhibits promising results in suppressing the consequences associated with HFD-induced obesity. It has the potential to be incorporated into food products benefiting consumer health and food industries.

Effects of dietary guava leaf aqueous extract supplementation on growth, antioxidant capacity, and non-specific immunity in mud crab Scylla paramamosain

Mud crab (Scylla paramamosain) fed five different diets with varying concentrations of guava leaf aqueous extract (0 mg·kg–1, 80 mg·kg–1, 160 mg·kg–1, 320 mg·kg–1, and 640 mg·kg–1) for 30 days. Mud crabs in the 320 mg·kg–1 guava-leaf extract groups outperformed the control group in terms of survival rates (SR), weight gain rates (WGR), and specific growth rates (SGR). When compared to the control group, mud crabs in the 320 mg·kg–1 guava-leaf extract groups had significantly higher levels of lipase (LPS), pepsin, lysozyme (LZM), superoxide dismutase (SOD), acid phosphatase (ACP), and glutathione (GSH) (P &lt; 0.05). The amylase (AMS) activity was significantly decreased in all experimental groups (P &lt; 0.05). Malondialdehyde (MDA) content in the hepatopancreas of mud crabs in the 160 mg·kg–1, 320 mg·kg–1, and 640 mg·kg–1 guava-leaf extract groups were significantly reduced compared to the control group (P &lt; 0.05). Additionally, real-time PCR results illustrated that the expression levels of GPx3, CAT, and JNK were all considerably increased in the 80 mg·kg–1 guava-leaf extract groups compared to the control group (P &lt; 0.05). In the 160 mg·kg–1, 320 mg·kg–1, and 320 mg·kg–1 guava-leaf extract groups, the expression levels of SOD genes were considerably greater than the control (P &lt; 0.05), which was consistent with the level of SOD activity. GST and P53 gene expression levels were significantly up-regulated in the 80 mg·kg–1, 160 mg·kg–1, 320 mg·kg–1, and 640 mg·kg–1 guava-leaf extract groups compared to the control group (P &lt; 0.05). Overall, the addition of 160 mg·kg–1-320 mg·kg–1 guava-leaf extract to the feed of Scylla paramamosain promoted growth, enhanced the activities of digestive and antioxidant enzymes, and strengthened immunity.

Validation of an online version of the Trier Social Stress Test in adult men and women

The Trier Social Stress Test (TSST) is a reliable laboratory protocol to induce acute psychosocial stress in the laboratory. If circumstances do not allow in-person assessments, an online version of the TSST could create more flexible research opportunities. To date, studies have confirmed subjective and autonomic stress responses to online TSST protocols. In this preregistered study, we focused on the effect of a TSST online for adults (TSST-OA) on cortisol and alpha amylase levels, and pleasure and arousal ratings. We hypothesized significant increases in cortisol, alpha amylase and arousal, and a decrease in pleasure in response to the TSST-OA. Also, we expected stronger cortisol responses in males as compared with females, as in the laboratory TSST. N=48 adults (56% female, meanage=23.02, SD=3.19) participated in the study. Saliva sampling devices were sent to participants’ home before testing sessions, during which the experimenter, a mixed-sex panel, and the participant joined a video call. Participants underwent the TSST-OA and overall provided five saliva samples for cortisol and alpha amylase detection. Pleasure and arousal ratings and psychometric questionnaires were also completed online. As hypothesized, the TSST-OA significantly increased cortisol, alpha amylase, and arousal levels, while it decreased pleasure. Moreover, cortisol responses were significantly stronger in males as compared to females. 64% of subjects were classified as responders (cortisol rise>1.5nmol/l). The TSST-OA successfully induced psychophysiological stress in adults. Our protocol offers new possibilities to study stress outside of the laboratory.

Context and Neuroendocrine Interplay in the Lab and Everyday Experiences: From Early Life to Adulthood

Context and Neuroendocrine Interplay in the Lab and Everyday Experiences: From Early Life to Adulthood

Indole 3-heterocyclic derivative: A potential antioxidant, antidiabetic agent and their docking study on alpha amylase

α-Amylase enzymes play a crucial role in breaking down starch molecules into simple sugar units that are easily absorbed into the bloodstream, resulting in a sudden increase in blood glucose levels. To respond this effect, there is a need for potent inhibitors that can hinder the enzymatic action of α-amylase. In this regard, indole-3-heterocycles (13−23) were synthesized via functionalizing of 3-(4-chlorobutyl)-1H-indole-5-carbonitrile with different triazoles, tetrazole, thiazole and mercapto benzothiazole with a base. Analytical techniques like 1H, 13C NMR, HRMS and IR spectroscopy characterized the newly synthesized indole heterocycles. These indole compounds functionalized were investigated for antidiabetic properties and antioxidant potentials on varying the side-chain. The synthesized indole compounds exhibited varying inhibition compared to α-amylase standard with IC50 values ranging between 6.44±1.14 μg/mL and 29.27±1.06 μg/mL competing with standard acarbose (IC50 = 5.7 ± 0.99 μg/mL). Among the series of eleven compounds, higher activity of IC50 = 6.44 ± 1.14 μg/mL was exhibited by compound 19 against α-amylase. In addition, an antioxidant potential of IC50 values of 15.62±1.20 μg/mL and 12.00 ± 1.00 μg/mL in 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and 2,2′-casino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays, respectively. The binding interactions of indole-3-heterocycles (13–23) were evaluated via in-silico docking performed on 5KEZ enzyme's active site. The current study has shown a light on the potential antidiabetic and antioxidant properties of lead molecules.

The association between cortisol-AA coordination in response to stress, negative urgency, and antisocial behavior in an urban adult sample

Cross-system coordination allows direct assessment of the degree of co-changing between systems within a person over a time course. Cross-system coordination of the HPA axis and the autonomic nervous system (ANS) in response to stress has implications for behavior problems. One previous study has identified that greater HPA axis-ANS coordination is linked to more behavior problems in youth. This study aims to extend the findings to adults and a new context, disadvantageous urban neighborhoods. Further, this study examined whether HPA axis-ANS coordination (indexed by cortisol and alpha amylase [AA], respectively) is associated with antisocial behavior and a conceptually relevant trait, negative urgency. A heterogeneous sample adults (n=124; college students, active offenders, and demographically matched controls) was recruited from a southern city. Self-report instruments assessed negative urgency and antisocial behavior. Participants completed a modified Trier Social Stress Test (mTSST), and provided four saliva samples before and after the mTSST to index cortisol and AA. Cortisol-AA stress coordination was stronger among offenders than other groups. Cortisol-AA stress coordination was positively associated with antisocial behavior and negative urgency. Findings support the interpretation that cortisol-AA stress coordination may indicate an overshooting ANS response at high emotional distress, highlight the importance of the multisystem approach in gaining new insights in behavior research, and extend prior work to a new context, disadvantageous urban neighborhoods. Future work is needed to understand whether early prolonged exposure to stressors in this context may alter individuals’ cross-system stress coordination and confer risk for antisocial behavior.

Objective Negative Evaluation Level, Perceived Stress, and Autonomic Reactivity in a Lab-based Stress Induction: A Test of Moderated Mediation in a Three-Variable System

Objective stress exposure (the impartial level of threat) and perceptions of stress (subjective level of threat a person endorses feeling) have both been closely linked to physiological and emotional stress-responding. The autonomic nervous system (ANS) is stress-responsive, integral to physical health, and implicated in mental health outcomes. We hypothesize that perceived stress is likely to be one mechanism (i.e., mediator) via which objective stress exerts an effect on ANS reactivity, but also the relationship between perceived stress and ANS reactivity may intensify as objective stress increases (i.e., moderation)—that is, potential moderated mediation in a 3-variable system (Goldstein et al., 2021). 128 emerging adults were randomized to three versions of the Trier Social Stress Test (TSST) that varied in objective level of negative evaluation: none (a control), intermediate/ambiguous, and explicit. The Primary Appraisal Secondary Appraisal Scale (PASA) measured perceived stress following TSST preparation. Salivary alpha amylase, systolic and diastolic blood pressure (SBP, DBP) and heart rate (HR) were measured four times. We tested the hypotheses of moderated mediation in a three-variable system with separate models for each outcome in Mplus Version 8. Objective stress (TSST Condition) directly predicted SBP (p=.018) but did not predict other ANS outcomes. Moreover, no significant moderated mediation (indirect effects varying on objective negative evaluation level) was observed. Stress condition was weakly related to both ANS reactivity and to our measure of perceived stress, Primary Appraisal from the PASA, and future work could investigate more sensitive mediators and outcomes.

Responses of soil labile organic carbon fractions and enzyme activities to long-term vegetation restorations in the karst ecosystems, Southwest China

To reveal the effects of karst long-term vegetation restorations on total soil organic carbon (TSOC), labile organic carbon fractions and enzymes, in the present study, the typical karst vegetation restoration region of Southwest China was selected as study area. Four types of long-term vegetation restoration engineering including seven kinds of planting measures were selected as study objects. Responses of TSOC and its labile fractions accumulation and distribution to the different vegetation restorations were studied. And the effects of these vegetation restorations on soil enzymes activities of urease, sucrase, amylase, and alkaline phosphatase were also researched. Results showed the vegetation restorations significantly improved TSOC accumulation and obviously changed microbial biomass carbon (MBC), easy oxidated organic carbon (EOC), and dissolved organic carbon (DOC) contents and their proportions to TSOC. And the effects of these different vegetation restorations on TSOC and its labile fractions were obviously different. TSOC content and reserves, MBC and EOC contents of Cupressus funebris and Lonicera japonica planting were high. While TSOC content and reserves of the two grassland restoration measures were the lowest. The vegetation restorations significantly increased soil enzyme activities of amylase (AMY), alkaline phosphatase (ALP), urease (URE) and sucrose (SUC). And there were significant differences in responses of different soil enzyme activities to vegetation restorations. The four soil enzyme activities were significantly correlated with TSOC and its labile fractions contents, especially soil AMY and ALP. Soil ALP and AMY were closely related to TSOC mineralization and accumulation and the formation and transformation of labile organic carbon fractions in Southwest China karst ecosystems. The present results provided scientific bases for the vegetation restoration of degraded karst ecosystems and were of great significance in carbon cycle intervention based on land use regulation in Southwest China.

The effects of hydrocortisone and yohimbine on human behavior in approach-avoidance conflicts

RationaleBalancing approach of positive and avoidance of negative stimuli is essential when faced with approach-avoidance conflicts, e.g., situations with both positive and negative outcomes. This balance is disturbed in several mental disorders, e.g., excessive avoidance in anxiety disorders, and heightened approach in substance use disorders. Since stress is assumed to impact these disorders’ etiology and maintenance, it seems crucial to understand how stress influences behavior in approach-avoidance conflicts. Indeed, some studies suggested altered approach-avoidance behavior under acute stress, but the mechanism underlying these effects is unknown.ObjectivesInvestigate how the pharmacological manipulation of major stress mediators (cortisol and noradrenaline) influences task-based approach-avoidance conflict behavior in healthy individuals.MethodsNinety-six participants (48 women, 48 men) received either 20mg hydrocortisone, 20mg yohimbine, both, or placebo before performing a task targeting foraging under predation in a fully crossed double-blind between-subject design. Moreover, we investigated effects of gender and endogenous testosterone and estradiol levels on approach-avoidance behavior.ResultsWhile biological stress markers (cortisol concentration, alpha amylase activity) indicated successful pharmacological manipulation, behavior in approach-avoidance conflicts was not affected as expected. Although yohimbine administration affected risky foraging latency under predation, we found no main effect of hydrocortisone or their interaction on behavior. In contrast, we found gender differences for almost all behavioral outcome measures, which might be explained by differences in endogenous testosterone levels.ConclusionsThe investigated major stress mediators were not sufficient to imitate previously shown stress effects on approach-avoidance conflict behavior. We discuss potential reasons for our findings and implications for future research.

Fabrication of protein crocin nanocomplex and its addition in proso millet flour for development of functional snacks with anti‐diabetic, anti‐hypertensive and antioxidant properties

SummaryThe current study was undertaken to develop novel gluten‐free functional snacks using proso millet flour with the addition of nano‐encapsulated crocin in sorghum (SPCS), pearl millet (PPCS) and foxtail millet (FPCS) protein nanoparticles. The crocin content was well retained (58.08%–77.67%) in the developed snacks. Characteristic properties like nutraceutical potential, rheological, sensory and physical attributes were evaluated. The extruded snacks showed good antioxidant (DNA damage inhibition assay), anti‐diabetic (alpha amylase inhibition assay) and anti‐hypertensive (angiotensin converting enzyme assay) properties. The developed snacks (P &gt; 0.05) showed higher elastic behaviour (G′) compared to viscous behaviour (G″). Hardness value for functional snacks were found to be in the range of 407–115 N. The control snacks showed significantly higher expansion ratio (348.76%) compared to functional snacks. The developed novel functional snacks appeared darker than control snacks as evaluated by colorimetric assay. The overall acceptability of the snacks was found to be equal on a 9‐point hedonic scale.

Molecular Characterization of spa, hld, fmhA, and lukD Genes and Computational Modeling the Multidrug Resistance of Staphylococcus Species through Callindra harrisii Silver Nanoparticles.

The problem of multidrug resistance in bacterial pathogens is significant and is related to the high morbidity and death rates of living things due to increased levels of beta-lactamases. Plant-derived nanoparticles have gained a great significance in the field of science and technology to combat bacterial diseases, especially multidrug-resistant bacteria. This study examines the multidrug resistance and virulent genes of identified pathogenic Staphylococcus species obtained from Molecular Biotechnology and Bioinformatics Laboratory (MBBL), culture collection. The polymerase chain reaction-based characterization of Staphylococcus aureus and Staphylococcus argenteus having ON875315.1 and ON876003.1 accession IDs revealed the presence of the spa, LukD, fmhA, and hld genes. The green synthesis of silver nanoparticles (AgNPs) was carried out by utilizing the leaf extract of Calliandra harrisii, of which metabolites act as capping and reducing agents for the precursor of nano-synthesis, i.e., AgNO3 of 0.25 M. The synthesized AgNPs were characterized via UV-vis spectroscopy, Fourier transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive X-ray analysis which inferred the bead-like shape of our nanoparticles with the size of 2.21 nm with the existence of aromatic and hydroxyl functional groups at surface plasmon resonance of 477 nm. The antimicrobial activity by AgNPs showed 20 mm inhibition of Staphylococcus species as compared to the vancomycin and cefoxitin antibiotics along with crude plant extract, which showed a minimum zone of inhibition. The synthesized AgNPs were also analyzed for various biological activities like anti-inflammatory with 99.15% inhibition in protein denaturation, antioxidant with 99.8% inhibition in free radical scavenging, antidiabetic with 90.56% inhibition of alpha amylase assay, and anti-haemolytic with 89.9% inhibition in cell lysis which shows good bioavailability and biocompatibility of the nanoparticles with the biological system of the living being. The amplified genes (spa, LukD, fmhA, and hld) were also analyzed for their interaction with AgNPs computationally at the molecular level. The 3-D structure of AgNP and amplified genes was retrieved from ChemSpider (ID: 22394) and Phyre2 online server, respectively. The binding affinities of AgNP with spa, LukD, fmhA, and hld were -7.16, -6.5, -6.45, and -3.3 kJ/mol, respectively, which infers a good docking score except of hld which is -3.3 kJ/mol due to its small size. The salient features of biosynthesized AgNPs proved to be an effective approach in combating the multidrug-resistant Staphylococcus species in the future.