Evaluation of 3-Deoxy-D-Arabino-Heptulosonate 7-Phosphate Synthase (DAHPS) as a Vulnerable Target in Mycobacterium tuberculosis.

Tuberculosis (TB) remains one of the leading causes of death due to a single pathogen. The emergence and proliferation of multidrug-resistant (MDR-TB) and extensively drug-resistant strains (XDR-TB) represent compelling reasons to invest in the pursuit of new anti-TB agents. The shikimate pathway, responsible for chorismate biosynthesis, which is a precursor of important aromatic compounds, is required for Mycobacterium tuberculosis growth. The enzyme 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase (MtbDAHPS) catalyzes the first step in the shikimate pathway and it is an attractive target for anti-tubercular agents. Here, we used a CRISPRi system to evaluate the DAHPS as a vulnerable target in M. tuberculosis. The silencing of aroG significantly reduces the M. tuberculosis growth in both rich medium and, especially, in infected murine macrophages. The supplementation with amino acids was only able to partially rescue the growth of bacilli, whereas the Aro supplement (aromix) was enough to sustain the bacterial growth at lower rates. This study shows that MtbDAHPS protein is vulnerable and, therefore, an attractive target to develop new anti-TB agents. In addition, the study contributes to a better understanding of the biosynthesis of aromatic compounds and the bacillus physiology. IMPORTANCE Determining the vulnerability of a potential target allows us to assess whether its partial inhibition will impact bacterial growth. Here, we evaluated the vulnerability of the enzyme 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase (DAHPS) from M. tuberculosis by silencing the DAHPS-coding aroG gene in different contexts. These results could lead to the development of novel and potent anti-tubercular agents in the near future.

Influence of l-carnitine on lipid metabolism of buffalo cumulus-oocyte complexes matured in either fetal bovine serum or fatty acid-free bovine serum albumin.

Consequences of oocyte supplementation with l-carnitine may vary depending on species-specific cellular lipid profile, level of mitochondrial activity, or even on ipid availability in culture medium. This study aimed to evaluate l-carnitine supplementation on competence and gene expression of enzymes related to lipid metabolism in oocytes and cumulus cells from buffalo COCs matured in the presence or absence of fetal bovine serum (FBS). COCs were matured in vitro in FBS (10%) or bovine serum albumin fatty acid-free (BSA-FAF) (0.4%) and with or without supplementation with l-carnitine (3.03 mM). COCs matured in the presence of FBS or BSA-FAF were fertilized and cultured, then supplemented with l-carnitine during in vitro maturation or in vitro embryo culture. Finally, in vivo mature and immature COCs were included for gene expression analysis. COCs matured in culture medium with FBS in the presence of l-carnitine produced a lower blastocyst rate (p ≤ 0.05) compared to controls. In turn, the blastocyst rate from COCs matured with BSA-FAF in the presence of l-carnitine was similar to controls (p > 0.05), and higher than FBS + L-carnitine treated COCs (p ≤ 0.05). Addition of l-carnitine during embryo culture showed no differences in blastocyst production between experimental groups and controls (p > 0.05). In cumulus cells, gene expression of ACACA, SCD and FASN was upregulated in COCs matured in the presence of BSA-FAF + L-carnitine, while all genes in oocytes were significantly expressed upregulated by COCs matured in vivo, and only BSA-FAF + L-carnitine group showed similar expression of the FASN gene. In conclusion, the consequences of l-carnitine supplementation during in vitro maturation of buffalo COCs on oocyte competence vary depending on presence or absence of FBS in culture. With FBS, l-carnitine impairs oocyte competence, while in its absence, gene expression suggests adequate lipid metabolism and increased oocyte competence.

The Effects of Fortetropin Supplementation on Body Composition, Strength, and Power in Humans and Mechanism of Action in a Rodent Model.

OBJECTIVE: The purpose of this study was to investigate the effects of Fortetropin on skeletal muscle growth and strength in resistance-trained individuals and to investigate the anabolic and catabolic signaling effects using human and rodent models. METHODS: In the rodent model, male Wistar rats (250 g) were gavage fed with either 1.2 ml of tap water control (CTL) or 0.26 g Fortetropin for 8 days. Then rats participated in a unilateral plantarflexion exercise bout. Nonexercised and exercised limbs were harvested at 180 minutes following and analyzed for gene and protein expression relative to mammalian target of rapamycin (mTOR) and ubiquitin signaling. For the human model, 45 (of whom 37 completed the study), resistance-trained college-aged males were divided equally into 3 groups receiving a placebo macronutrient matched control, 6.6 or 19.8 g of Fortetropin supplementation during 12 weeks of resistance training. Lean mass, muscle thickness, and lower and upper body strength were measured before and after 12 weeks of training. RESULTS: The human study results indicated a Group × Time effect (p ≤ 0.05) for lean mass in which the 6.6 g (+1.7 kg) and 19.8 g (+1.68 kg) but not placebo (+0.6 kg) groups increased lean mass. Similarly, there was a Group × Time effect for muscle thickness (p ≤ 0.05), which increased in the experimental groups only. All groups increased equally in bench press and leg press strength. In the rodent model, a main effect for exercise (p ≤ 0.05) in which the control plus exercise but not Fortetropin plus exercise increased both ubiquitin monomer protein expression and polyubiquitination. mTOR signaling was elevated to a greater extent in the Fortetropin exercising conditions as indicated by greater phosphorylation status of 4EBP1, rp6, and p70S6K for both exercising conditions. CONCLUSIONS: Fortetropin supplementation increases lean body mass (LBM) and decreases markers of protein breakdown while simultaneously increasing mTOR signaling.

Effects of Arachidonic Acid Supplementation on Acute Anabolic Signaling and Chronic Functional Performance and Body Composition Adaptations.

BACKGROUND: The primary purpose of this investigation was to examine the effects of arachidonic acid (ARA) supplementation on functional performance and body composition in trained males. In addition, we performed a secondary study looking at molecular responses of ARA supplementation following an acute exercise bout in rodents. METHODS: Thirty strength-trained males (age: 20.4 ± 2.1 yrs) were randomly divided into two groups: ARA or placebo (i.e. CTL). Then, both groups underwent an 8-week, 3-day per week, non-periodized training protocol. Quadriceps muscle thickness, whole-body composition scan (DEXA), muscle strength, and power were assessed at baseline and post-test. In the rodent model, male Wistar rats (~250 g, ~8 weeks old) were pre-fed with either ARA or water (CTL) for 8 days and were fed the final dose of ARA prior to being acutely strength trained via electrical stimulation on unilateral plantar flexions. A mixed muscle sample was removed from the exercised and non-exercised leg 3 hours post-exercise. RESULTS: Lean body mass (2.9%, p<0.0005), upper-body strength (8.7%, p<0.0001), and peak power (12.7%, p<0.0001) increased only in the ARA group. For the animal trial, GSK-ß (Ser9) phosphorylation (p<0.001) independent of exercise and AMPK phosphorylation after exercise (p-AMPK less in ARA, p = 0.041) were different in ARA-fed versus CTL rats. CONCLUSIONS: Our findings suggest that ARA supplementation can positively augment strength-training induced adaptations in resistance-trained males. However, chronic studies at the molecular level are required to further elucidate how ARA combined with strength training affect muscle adaptation.

Effects of chronic Bryophyllum pinnatum administration on Wistar rat pregnancy.

BACKGROUND: Drugs used in preterm labor (PTL) have side effects. Research into new tocolytic agents is recommended. The plant Bryophyllum pinnatum (Lam.) Oken (Kalanchoe pinnata (Lam.) Pers) is used in Anthroposophic Medicine for PTL, insomnia, and emotional disturbances. The aim of this study was to evaluate the effects of B. pinnatum mother tincture (MT) on Wistar rats and their fetuses throughout pregnancy. METHOD: Sixty animals were divided into 6 equal groups: controls C1 and C2 received 1 and 25 × the maximum daily dose of a 30% ethanol / water solution, serving as vehicle of B. pinnatum MT; B1 and B2 received 1 and 25 × the maximum daily dose of B. pinnatum MT; and B3 and B4 received 50 and 100 × B. pinnatum concentrate. The following parameters were assessed: weight gain; maternal and fetal mortality; implantations and resorptions; number and weight of fetuses and placentas; major external fetal malformations. RESULTS: Rat weight gain (excluding fetal and placental weight) was higher in group B4 and lower in group B2. There were no maternal or fetal deaths and no group differences in implantations and resorptions or number and weight of fetuses and placentas. No macroscopic fetal abnormalities were observed at the 4 dosage levels investigated. CONCLUSION: Daily administration of B. pinnatum MT at high doses to pregnant Wistar rats interfered with maternal weight gain and did not interfere with fetal compartment.

Effects of 8 weeks of Xpand® 2X pre workout supplementation on skeletal muscle hypertrophy, lean body mass, and strength in resistance trained males.

BACKGROUND: Xpand® 2X is a proprietary blend comprised of branched chain amino acids, creatine monohydrate, beta-alanine (CarnoSyn®), quercetin, coenzymated B-vitamins, alanyl-glutamine (Sustamine®), and natural nitrate sources from pomegranate and beet root extracts purported to enhance the neuromuscular adaptations of resistance training. However to date, no long-term studies have been conducted with this supplement. The purpose of this study was to investigate the effects of a multi-ingredient performance supplement (MIPS) on skeletal muscle hypertrophy, lean body mass and lower body strength in resistance-trained males. METHODS: Twenty resistance-trained males (21.3 ± 1.9 years) were randomly assigned to consume a MIPS or a placebo of equal weight and volume (food-grade orange flavors and sweeteners) in a double-blind manner, 30 minutes prior to exercise. All subjects participated in an 8-week, 3-day per week, periodized, resistance-training program that was split-focused on multi-joint movements such as leg press, bench press, and bent-over rows. Ultrasonography measured muscle thickness of the quadriceps, dual-energy X-ray absorptiometry (DEXA) determined lean body mass, and strength of the bench press and leg press were determined at weeks 0, 4, and 8 of the study. Data were analyzed with a 2 × 3 repeated measures ANOVA with LSD post hoc tests utilized to locate differences. RESULTS: There was a significant group-by-time interaction in which the MIPS supplementation resulted in a significant (p < 0.01) increase in strength of the bench press (18.4% vs. 9.6%) compared with placebo after 4 and 8 weeks of training. There were no significant group by time interactions between MIPS supplementation nor the placebo in leg press strength (p = .08). MIPS supplementation also resulted in a significant increase in lean body mass (7.8% vs. 3.6%) and quadriceps muscle thickness (11.8% vs. 4.5%) compared with placebo (group*time, p <0.01). CONCLUSIONS: These results suggest that this MIPS can positively augment adaptations in strength, and skeletal muscle hypertrophy in resistance-trained men.

The effects of 8 weeks of whey or rice protein supplementation on body composition and exercise performance.

Consumption of moderate amounts of animal-derived protein has been shown to differently influence skeletal muscle hypertrophy during resistance training when compared with nitrogenous and isoenergetic amounts of plant-based protein administered in small to moderate doses. Therefore, the purpose of the study was to determine if the post-exercise consumption of rice protein isolate could increase recovery and elicit adequate changes in body composition compared to equally dosed whey protein isolate if given in large, isocaloric doses.

[The socio-cultural bases of alternative therapeutic practices]. / Bases socioculturais das práticas terapêuticas alternativas.

The emergence and development of alternative therapies comprised part of the counter-cultural movement initiated in the 1960s. In the health field of the western world, the social transformations that occurred at that time inaugurated a period in which diverse health cultures coexisted. In this work we elaborate the socio-anthropological interpretations of cultural, political and socio-economic factors that influenced these transformations. First, we identify the macrocultural factors that would influence the transformations in the health field. Next, within this field we analyze the conflicts and disruptions that contributed to the search for new therapeutic practices. We use text analysis as the principal methodology, starting from the presupposition that the integrative approach of alternative therapies expresses an aspect of the transformation in the cultural values contemporaneous societies.

Meditation in health: an operational definition.

Despite its evergrowing use in health-related areas, procedures characterized as meditation have been little or not at all defined operationally, which hinders its use in a standardized manner. In the present study, the authors present a possible operational definition of meditation, which has been used in social and academic projects, developed in Universidade Federal de São Paulo. In this proposal, it is emphasized that, in order to be characterized as meditation, the procedure should encompass the following requirements: (1) the use of a specific technique (clearly defined), (2) muscle relaxation in some moment of the process and (3) "logic relaxation"; (4) it must necessarily be a self-induced state, and (5) use of "self-focus" skill (coined "anchor").