Involvement of Serine Threonine Protein Kinase, PknL, from Mycobacterium Tuberculosis H37Rv in Starvation Response of Mycobacteria

The adaptation to nutrient depletion in bacteria involves a highly organized series of intracellular events that enable them to adapt to starvation conditions. The regulatory effect of serine threonine protein kinase, PknL, from Mycobacterium tuberculosis strain H37Rv was investigated under nutrient deprived conditions that simulate circumstances leading to latency. Recombinant PknL was expressed in Mycobacterium smegmatis strain mc2155 in its wild type and mutant forms. In vitro growth kinetics experiments revealed that clone expressing active PknL had a significant growth advantage under nutrient limiting conditions. Experiments were conducted to ascertain the in silico predictions of the involvement of PknL in regulating glutamine metabolism in mycobacteria. Furthermore, a role for PknL in cell wall biogenesis/cell division was shown by scanning electron microscopy

One-year dietary supplementation with walnuts modifies exosomal miRNA in elderly subjects

Purpose: Epidemiological studies and clinical trials support the association of nut consumption with a lower risk of prevalent non-communicable diseases, particularly cardiovascular disease. However, the molecular mechanisms underlying nut benefits remain to be fully described. MicroRNAs (miRNAs) are post-transcriptional regulators of gene expression and play a pivotal role in health and disease. Exosomes are extracellular vesicles released from cells and mediate intercellular communication. Whether nut consumption modulates circulating miRNAs (c-miRNAs) transported in exosomes is poorly described. Methods: Cognitively healthy elderly subjects were randomized to either control (n = 110, abstaining from walnuts) or daily supplementation with walnuts (15% of their total energy, ≈30–60 g/day, n = 101) for 1-year. C-miRNAs were screened in exosomes isolated from 10 samples, before and after supplementation, and identified c-miRNA candidates were validated in the whole cohort. In addition, nanoparticle tracking analysis and lipidomics were assessed in pooled exosomes from the whole cohort. Results: Exosomal hsa-miR-32-5p and hsa-miR-29b-3p were consistently induced by walnut consumption. No major changes in exosomal lipids, nanoparticle concentration or size were found. Conclusion: Our results provide novel evidence that certain c-miRNAs transported in exosomes are modulated by walnut consumption. The extent to which this finding contributes to the benefits of walnuts deserves further research.Fundacion Ramon Areces (CIVP18A3888) Madrid, Spain; the Instituto de Salud Carlos III-Fondo de Investigacion Sanitaria–Fondo Europeo de Desarrollo Regional (grant PI15/01014 and PI18/01152), and the Spanish Agencia Estatal de Investigacion and European Feder Funds (AGL2016-78922-R, PID2019-109369RB-I00, RTI2018-093873-A-I00 and BIO2017-86500-R). AS-V is recipient of the Instituto de Salud Carlos III Miguel Servet II fellowship (grant CP II 17/00029

The Walnuts and Healthy Aging study (WAHA): Protocol for a Nutritional Intervention Trial with Walnuts on Brain Aging

Introduction: An unwanted consequence of population aging is the growing number of elderly at risk of neurodegenerative disorders, including dementia and macular degeneration. As nutritional and behavioral changes can delay disease progression, we designed the Walnuts and Healthy Aging (WAHA) study, a two-center, randomized, 2-year clinical trial conducted in free-living, cognitively healthy elderly men and women. Our interest in exploring the role of walnuts in maintaining cognitive and retinal health is based on extensive evidence supporting their cardio-protective and vascular health effects, which are linked to bioactive components, such as n-3 fatty acids and polyphenols. Methods: The primary aim of WAHA is to examine the effects of ingesting walnuts daily for 2 years on cognitive function and retinal health, assessed with a battery of neuropsychological tests and optical coherence tomography, respectively. All participants followed their habitual diet, adding walnuts at 15% of energy (≈30-60 g/day) (walnut group) or abstaining from walnuts (control group). Secondary outcomes include changes in adiposity, blood pressure, and serum and urinary biomarkers in all participants and brain magnetic resonance imaging in a subset. Results: From May 2012 to May 2014, 708 participants (mean age 69 years, 68% women) were randomized. The study ended in May 2016 with a 90% retention rate. Discussion: The results of WAHA might provide high-level evidence of the benefit of regular walnut consumption in delaying the onset of age-related cognitive impairment and retinal pathology. The findings should translate into public health policy and sound recommendations to the general population

Moderate exercise-induced changes in the iron status of young adult women

The effect of moderate aerobic exercise on the iron status of previously sedentary college-age women was determined by conducting a short-term (12 Wks) and a long-term (24 Wks) study. In the first study, subjects (N = 60) were randomly assigned to one of the following groups: (1) 50 mg/d iron supplement, low iron diet; (2) 10 mg/d iron supplement, low iron diet; (3) Placebo, free choice diet; (4) Meat supplement to achieve recommended allowance for iron; and (5) Control, free choice diet. The long-term study (N = 62) was designed similarly and had all the above groups except the group that received the 10 mg/d iron supplement. In both the studies, all groups except the control were required to exercise a minimum of 3d/wk at 70-90% of their maximal heart rate. VO\sb{2max} was measured at baseline and at the end of the study. Blood was sampled at baseline and every 4 weeks thereafter. Iron status indices were measured in both the studies. Mechanisms inducing changes in the iron status were also elucidated in the long-term study. In the short-term study, a decline in hemoglobin at the end of 4 weeks was observed in all the exercising groups. Transferrin saturation and serum ferritin were also negatively impaired in exercising subjects in study 1. The results from the long-term study in terms of changes in iron status were not as striking as the short-term study probably because of a higher baseline iron status in these subjects. However, a repeated time measures analysis of covariance with baseline hemoglobin as covariate indicated a significantly (p $\le$ 0.05) lower post hemoglobin in the Placebo group of study 2, compared to the iron supplemented groups. A significant drop in serum ferritin was detected in the Placebo group after 24 weeks of exercise. The results from both these studies together indicate that moderate aerobic exercise can compromise iron status in previously untrained women. An initial declining trend in haptoglobins in the exercising groups and a significant increase in serum erythropoietin in the Placebo group after 24 weeks suggest that a mild hemolysis might be occurring with exercise. However, a significant increase in haptoglobins at week 24 in the Meat group and an overall improvement in the iron status indices in the meat and iron supplemented groups in both the studies indicate that the initial alterations in the iron status induced by moderate aerobic exercise can be offset by oral iron therapy or by meeting the daily iron requirements through muscle food consumption

Chromium(III)-induced apoptosis of lymphocytes: death decision by ROS and Src-family tyrosine kinases

Apoptosis is an active process induced by a variety of physiological and external stimuli, in which elimination of damaged cells are effected through a genetically controlled process. In this study, we have examined the mechanism of chromium(III) [Cr(III)]-induced cytotoxicity with respect to its relationship to oxidative stress. Morphology, flow cytometry, and DNA fragmentation studies show that tris-(1,10-phenanthroline)chromium(III) [Cr(III)-phen], tris-(2,2'-bipyridyl)chromium(III) [Cr(III)-bpy], trans-diaqua[1,2-bis(salicylideneamino)ethanechromium(III)] [Cr(III)-salen], and trans-diaqua[1,3-bis(salicylideneamino)propanechromium(III)] [Cr(III)-salprn] induced apoptosis of lymphocytes. Pentaammineaquachromium(III) [Cr(III)-hpa] does not induce apoptosis. Apoptosis induced by these complexes involves the generation of reactive oxygen species (ROS) as seen by increased fluorescence of dichloroflourescein (DCF) observed through flow cytometry. Pretreatment of lymphocytes with antioxidants completely abrogate apoptosis. Cr(III) treatment also increased the expression and activation of Src-family tyrosine kinases viz. p56lck, p59fyn, and p53/56lyn, as seen by immunoblotting and immune complex kinase assay. PP2, a selective Src-family tyrosine kinase inhibitor, abolishes apoptosis, indicating that Src-family tyrosine kinases are directly involved in eliciting apoptosis. Interestingly, a one-to-one correlation between the expression of Src-family tyrosine kinases and ROS is observed, since antioxidants pretreatment inhibits the expression and the activation of these kinases. These results further indicate that Cr(III)-induced apoptosis is mediated through production of ROS, which in turn activates the Src-family tyrosine kinases. The increased activation of Src-family tyrosine kinases may be a mechanism involved in apoptosis of lymphocytes elicited by various other physiological stimuli that exploit ROS as a second messenger

Effect of Altering Dietary n-6:n-3 Polyunsaturated Fatty Acid Ratio with Plant and Marine-Based Supplement on Biomarkers of Bone Turnover in Healthy Adults

Although there is accumulating evidence for a protective role of n-3 polyunsaturated fatty acids (n-3 PUFAs) on bone health, there are limited studies that examine the effect of altering dietary n-6:n-3 PUFA ratio with plant and marine sources of n-3 PUFA on bone health. Healthy adults (n = 24) were randomized into an eight-week crossover study with a four-week washout between treatments, with each subject consuming three of four diets. The four diets differed in the dietary n-6:n-3 PUFA ratios and either had an algal oil supplement added or not: (Control diet (10:1); α-linolenic acid (ALA) diet (2:1); Eicosapentaenoic acid/Docosahexaenoic acid (EPA/DHA) diet (10:1 plus supplement (S) containing EPA/DHA; Combination diet (2:1 + S)). The supplement was microalgae oil that provided 1 g EPA + DHA/day. Flaxseed oil and walnuts provided 8.6 g of ALA/day in the 2:1 diets. Serum levels of c-telopeptide (CTX), procollagen Type I N-terminal peptide, and osteocalcin showed significant correlation with age but none of the bone markers or peroxisomal proliferator-activated receptor-γ mRNA expression was significantly different between the diets. Serum CTX was negatively associated with red blood cell membrane linoleic acid and ALA and positively associated with membrane DHA. Neither altering dietary n-6:n-3 PUFA ratio from a 10:1 to a 2:1 ratio nor adding EPA/DHA supplement significantly changed bone turnover in the short term in healthy adults

Evaluation of a Validated Food Frequency Questionnaire for Self-Defined Vegans in the United States

This study aimed to develop and validate a de novo food frequency questionnaire for self-defined vegans in the United States. Diet histories from pilot samples of vegans and a modified ‘Block Method’ using seven selected nutrients of concern in vegan diet patterns, were employed to generate the questionnaire food list. Food frequency responses of 100 vegans from 19 different U.S. states were obtained via completed mailed questionnaires and compared to multiple telephone-conducted diet recall interviews. Computerized diet analyses were performed. Correlation coefficients, t-tests, rank, cross-tabulations, and probability tests were used to validate and compare intake estimates and dietary reference intake (DRI) assessment trends between the two methods. A 369-item vegan-specific questionnaire was developed with 252 listed food frequency items. Calorie-adjusted correlation coefficients ranged from r = 0.374 to 0.600 (p < 0.001) for all analyzed nutrients except calcium. Estimates, ranks, trends and higher-level participant percentile placements for Vitamin B12 were similar with both methods. Questionnaire intakes were higher than recalls for most other nutrients. Both methods demonstrated similar trends in DRI adequacy assessment (e.g., significantly inadequate vitamin D intake among vegans). This vegan-specific questionnaire can be a useful assessment tool for health screening initiatives in U.S. vegan communities

Dietary Animal to Plant Protein Ratio Is Associated with Risk Factors of Metabolic Syndrome in Participants of the AHS-2 Calibration Study

Background: Few research studies have focused on the effects of dietary protein on metabolic syndrome and its components. Our objective was to determine the relationship between the type of dietary protein intake and animal to plant (AP) protein ratio with metabolic syndrome and its components. Methods: This population-based study had a cross sectional design and conducted on 518 participants of the Adventist Health Study 2 (AHS-2) Calibration Study. Two sets of three dietary 24-h recalls were obtained six months apart. Anthropometric measures and biochemical tests were performed in clinics. Regression calibration models were used to determine the association of type of dietary protein with metabolic syndrome and its components (raised triglyceride, raised blood pressure, reduced high-density lipoprotein cholesterol (HDL), raised fasting blood glucose and increased waist circumference). Results: The likelihood of metabolic syndrome was lower in those with higher total dietary protein and animal protein intake (p = 0.02).Total protein (β = 0.004, [95%CI: 0.002, 0.007]), animal protein intake (β = 0.004, [95%CI: 0.001, 0.007]) and AP protein intake ratio (β = 0.034, [95%CI: 0.021, 0.047]) were positively associated with waist circumference. Higher AP protein ratio was related to higher fasting blood glucose (β = 0.023, [95%CI: 0.005, 0.041]). Conclusion: Our study suggests that considering a significant amount of plant protein as a part of total dietary protein has beneficial effects on cardiometabolic risk factors