Effects of Exercise Training and IL-6 Receptor Blockade on Gastric Emptying and GLP-1 Secretion in Obese Humans: Secondary Analyses From a Double Blind Randomized Clinical Trial.

BACKGROUND: Interleukin-6 (IL-6) is released from skeletal muscle during exercise and systemic IL-6 levels therefore increase acutely in response to a single bout of exercise. We recently showed that an acute increase in IL-6 delayed gastric emptying rate and improved postprandial glycemia. Here we investigate whether repeated increases in IL-6, induced by exercise training, influence gastric emptying rate and moreover if IL-6 is required for exercise-induced adaptations in glycemic control including secretion of glucagon and glucagon-like peptide-1 (GLP-1). METHODS: A total of 52 abdominally obese non-diabetic men and women were randomly assigned into four groups performing 12 weeks of endurance exercise or no exercise with or without IL-6 receptor blockade (tocilizumab). The primary endpoint was change in gastric emptying rate in response to the intervention and other endpoints included changes in glycemic control, glucagon, and GLP-1 secretion. RESULTS: There was no change in gastric emptying rate in any of the four groups following the intervention and comparing differences in change between groups also revealed no difference. Postprandial glucose remained unchanged in all groups but the exercise + tocilizumab group, which improved postprandial glucose in response to the intervention. The area under the curve for meal-stimulated glucagon, active and total GLP-1 increased in response to IL-6 receptor blockade, this effect was independent of exercise. CONCLUSION: Exercise training and long-term IL-6 receptor blockade did not change gastric emptying rates in obese humans. IL-6 receptor blockade increased glucagon and GLP-1 secretion and implicate IL-6 in the regulation of the human alpha and L cells.

Effect of Aerobic and Resistance Exercise on Cardiac Adipose Tissues: Secondary Analyses From a Randomized Clinical Trial.

Importance: Epicardial and pericardial adipose tissues are emerging as important risk factors for cardiovascular disease, and there is a growing interest in discovering strategies to reduce the accumulation of fat in these depots. Objective: To investigate whether a 12-week endurance or resistance training intervention regulates epicardial and pericardial adipose tissue mass. Design, Setting, and Participants: Secondary analysis of a randomized, assessor-blinded clinical trial initiated on August 2016 and completed April 2018. This single-center, community-based study included 50 physically inactive participants with abdominal obesity. Interventions: Participants were randomized to a supervised high-intensity interval endurance training (3 times a week for 45 minutes), resistance training (3 times a week for 45 minutes), or no exercise (control group). Main Outcomes and Measures: Change in epicardial and pericardial adipose tissue mass assessed by magnetic resonance imaging, based on a prespecified secondary analysis plan including 3 of 5 parallel groups. Results: Of 50 participants (mean [SD] age, 41 [14] years, 10 men [26%]; mean [SD] body mass index [calculated as weight in kilograms divided by height in meters squared], 32 [5]), 39 [78%] completed the study. Endurance training and resistance training reduced epicardial adipose tissue mass by 32% (95% CI, 10%-53%) and 24% (95% CI, 1%-46%), respectively, compared with the no exercise control group (56% [95% CI, 24%-88%]; P = .001 and 48% [95% CI, 15%-81%]; P < .001, respectively). While there was a nonsignificant reduction in pericardial adipose tissue mass after endurance training (11% [95% CI, -5% to 27%]; P = .17), resistance training significantly reduced pericardial adipose tissue mass by 31% (95% CI, 16%-47%; P < .001) when compared with the no exercise control group. Compared with the no exercise control group, there was an increase in left ventricular mass by endurance (20 g [95% CI, 11%-30%]; P < .001) and resistance training (18 g [95% CI, 8%-28%]; P < .001). Other cardiometabolic outcomes remained unchanged after the 12-week trial period. Conclusions and Relevance: In individuals with abdominal obesity, both endurance and resistance training reduced epicardial adipose tissue mass, while only resistance training reduced pericardial adipose tissue mass. These data highlight the potential preventive importance of different exercise modalities as means to reduce cardiac fat in individuals with abdominal obesity. Trial Registration: ClinicalTrials.gov identifier: NCT02901496.

Exercise-Induced Changes in Visceral Adipose Tissue Mass Are Regulated by IL-6 Signaling: A Randomized Controlled Trial.

Visceral adipose tissue is harmful to metabolic health. Exercise training reduces visceral adipose tissue mass, but the underlying mechanisms are not known. Interleukin-6 (IL-6) stimulates lipolysis and is released from skeletal muscle during exercise. We hypothesized that exercise-induced reductions in visceral adipose tissue mass are mediated by IL-6. In this randomized placebo-controlled trial, we assigned abdominally obese adults to tocilizumab (IL-6 receptor antibody) or placebo during a 12-week intervention with either bicycle exercise or no exercise. While exercise reduced visceral adipose tissue mass, this effect of exercise was abolished in the presence of IL-6 blockade. Changes in body weight and total adipose tissue mass showed similar tendencies, whereas lean body mass did not differ between groups. Also, IL-6 blockade increased cholesterol levels, an effect not reversed by exercise. Thus, IL-6 is required for exercise to reduce visceral adipose tissue mass and emphasizes a potentially important metabolic consequence of IL-6 blockade.

The role of exercise combined with tocilizumab in visceral and epicardial adipose tissue and gastric emptying rate in abdominally obese participants: protocol for a randomised controlled trial.

BACKGROUND: Exercise reduces the amount of visceral adipose tissue (VAT) and the risk of cardiometabolic diseases. The underlying mechanisms responsible for these exercise-induced adaptations are unclear, but they may involve lipolytic actions of interleukin-6 (IL-6). Contracting skeletal muscles secrete IL-6, leading to increased circulating IL-6 levels in response to exercise. The aim of this study is to investigate whether IL-6 is involved in mediating the effects of exercise on visceral and epicardial adipose tissue volume and glycaemic control. METHODS/DESIGN: Seventy-five physically inactive males and females aged > 18 years with a waist-to-height ratio > 0.5 and/or waist circumference ≥ 88 cm (females) or ≥ 102 cm (males) are being recruited to participate in a 12-week intervention study. Participants are randomly allocated to one of five groups (1:1:1:1:1). Two groups consist of supervised endurance exercise training combined with the IL-6 blocker tocilizumab (ET) or saline used as placebo (EP), two groups consist of no exercise combined with tocilizumab (NT) or placebo (NP), and one group consists of resistance exercise and placebo (RP). Although the study is an exploratory trial, the primary outcome is change in VAT volume from before to after intervention, with secondary outcomes being changes in (1) epicardial adipose tissue, (2) pericardial adipose tissue and (3) gastric emptying. Depots of adipose tissue are quantitated by magnetic resonance imaging Gastric emptying and glucose metabolism are assessed using mixed-meal tolerance tests. DISCUSSION: Understanding the role of IL-6 in mediating the effects of exercise on visceral and epicardial adipose tissue and glycaemic control may lead to novel therapeutic approaches in the prevention of cardiometabolic diseases. TRIAL REGISTRATION: ClinicalTrials.gov, NCT02901496 . Registered on 1 August 2016 and posted retrospectively on 15 September 2016.

Diversity of culturable bacteria recovered from Pico Bolívar's glacial and subglacial environments, at 4950 m, in Venezuelan tropical Andes.

Even though tropical glaciers are retreating rapidly and many will disappear in the next few years, their microbial diversity remains to be studied in depth. In this paper we report on the biodiversity of the culturable fraction of bacteria colonizing Pico Bolívar's glacier ice and subglacial meltwaters, at ∼4950 m in the Venezuelan Andean Mountains. Microbial cells of diverse morphologies and exhibiting uncompromised membranes were present at densities ranging from 1.5 × 104 to 4.7 × 104 cells/mL in glacier ice and from 4.1 × 105 to 9.6 × 105 cells/mL in subglacial meltwater. Of 89 pure isolates recovered from the samples, the majority were eurypsychrophilic or stenopsychrophilic, according to their temperature range of growth. Following analysis of their 16S rDNA nucleotidic sequence, 54 pure isolates were assigned to 23 phylotypes distributed within 4 different phyla or classes: Beta- and Gammaproteobacteria, Actinobacteria, and Bacteroidetes. Actinobacteria dominated the culturable fraction of glacier ice samples, whereas Proteobacteria were dominant in subglacial meltwater samples. Chloramphenicol and ampicillin resistance was exhibited by 73.07% and 65.38%, respectively, of the subglacial isolates, and nearly 35% of them were multiresistant. Considering the fast rate at which tropical glaciers are melting, this study confirms the urgent need to study the microbial communities immured in such environments.

Bioprospecting glacial ice for plant growth promoting bacteria.

Glaciers harbor a wide diversity of microorganisms, metabolically versatile, highly tolerant to multiple environmental stresses and potentially useful for biotechnological purposes. Among these, we hypothesized the presence of bacteria able to exhibit well-known plant growth promoting traits (PGP). These kinds of bacteria have been employed for the development of commercial biofertilizers; unfortunately, these biotechnological products have proven ineffective in colder climates, like the ones prevailing in mountainous ecosystems. In the present work, we prospected glacial ice collected from two small tropical glaciers, located above 4.900 m in the Venezuelan Andes, for cold-active PGP bacteria. The initial screening strategy allowed us to detect the best inorganic-P solubilizers at low temperatures, from a sub-sample of 50 bacterial isolates. Solubilization of tricalcium phosphate, aluminum- and iron-phosphate, occurred in liquid cultures at low temperatures and was dependent on medium acidification by gluconic acid production, when bacteria were supplied with an appropriate source of carbon. Besides, the isolates were psychrophilic and in some cases exhibited a broad range of growth-temperatures, from 4 °C to 30 °C. Additional PGP abilities, including phytohormone- and HCN production, siderophore excretion and inhibition of phytopathogens, were confirmed in vitro. Nucleotidic sequence analysis of 16S rRNA genes allowed us to place the isolates within the Pseudomonas genus. Our results support the possible use of these strains to develop cold-active biofertilizers to be used in mountainous agriculture.

Bacteria recovered from a high-altitude, tropical glacier in Venezuelan Andes.

Glacial-ice microorganisms are intensively studied world-wide for a number of reasons, including their psychrophilic lifestyle, their usefulness in biotechnology procedures and their relationship with the search of life outside our planet. However, because of the difficulties for accessing and working at altitudes of >5.000 m above sea level, tropical glaciers have received much less attention than their arctic and antarctic counterparts. In the present work we isolated and characterized a total of forty-five pure isolates originating from direct plating of melted ice collected at the base of a rapidly-retreating, small glacier located at around 4.900 m.a.s.l. in Mount Humboldt (Sierra Nevada National Park, Mérida State, Venezuela). Initial examination of melted ice showed the presence of abundant- (>106 cells ml⁻¹), morphologically diverse- and active bacterial cells, many of which were very small ("dwarf cells"). The majority of the isolates were psychrophilic or psychrotolerant and many produced and excreted cold-active extracellular enzymes (proteases and amylases). The antibiotic tests showed an elevated percentage of isolates resistant to high doses (100 µg/ml) of different antibiotics including ampicillin, penicillin, nalidixic acid, streptomycin, chloramphenicol, kanamycin and tetracycline. Multiresistance was also observed, with 22.22 % of the strains simultaneously resistant up to five of the antibiotics tested. Metal resistance against Ni⁺⁺, Zn⁺⁺ and Cu⁺⁺ was also detected. In accordance with these results, plasmids of low and high molecular weight were detected in 47 % of the isolates. Twenty-two partial 16S rDNA sequences analyzed allowed grouping the isolates within five different phyla/classes: Alpha-, Beta- and Gamma-proteobacteria, Actinobacteria and Flavobacteria. This is the first report concerning South American Andean glacial ice microorganisms.

Characterization of the mineral phosphate-solubilizing activity of Pantoea agglomerans MMB051 isolated from an iron-rich soil in southeastern Venezuela (Bolívar State).

The mineral phosphate-solubilizing (MPS) activity of a Pantoea agglomerans strain, namely MMB051, isolated from an iron-rich, acidic soil near Ciudad Piar (Bolívar State, Venezuela), was characterized on a chemically defined medium (NBRIP). Various insoluble inorganic phosphates, including tri-calcium phosphate [Ca(3)(PO(4))(2)], iron phosphate (FePO(4)), aluminum phosphate (AlPO(4)), and Rock Phosphate (RP) were tested as sole sources of P for bacterial growth. Solubilization of Ca(3)(PO(4))(2) was very efficient and depended on acidification of the external milieu when MMB051 cells were grown in the presence of glucose. This was also the case when RP was used as the sole P source. On the other hand, the solubilization efficiency toward more insoluble mineral phosphates (FePO(4) and AlPO(4)) was shown to be very low. Even though gluconic acid (GA) was detected on culture supernatants of strain MMB051, a consequence of the direct oxidation pathway of glucose, inorganic-P solubilization seemed also to be related to other processes dependent on active cell growth. Among these, proton release by ammonium (NH(4)(+) ) fixation appeared to be of paramount importance to explain inorganic-P solubilization mediated by strain MMB051. On the contrary, the presence of nitrate (NO(3)(-) ) salts as the sole N source affected negatively the ability of MMB051 cells to solubilize inorganic P.

Agricultural intensity and landscape structure: influences on the macroinvertebrate assemblages of small streams in northern Germany.

The present study aimed to relate aquatic macroinvertebrate community composition to agricultural intensity and landscape structure. A total of 360 streams were investigated within the Aller river basin in northern Germany. The study area is typical of central German arable agricultural regions, but the small streams were of low dilution potential. These streams were characterized for abiotic parameters (including modeled potential for diffuse inputs from agricultural sources) and macroinvertebrate communities, with data collected over a 17-year period. Spray drift potential did not correlate with community composition. In contrast, the relative index of runoff potential (RP) was negatively correlated with various measures of taxonomic richness and abundance. Community composition also was correlated with environmental parameters, including stream width, clay content of sediment, and presence of dead wood in sediment. The abundance of sensitive species decreased significantly during the main period of agrochemical use at sites of high RP but completely recovered by the following spring. Long-term decreased taxonomic richness and a shift to ecologically robust species also were observed at sites of high RP. The results suggest that long-term alterations in community measures probably were associated with factors related to runoff input. Nevertheless, the community composition remained reasonably rich and even. Landscape structure also appeared to influence community structure. Abundance of sensitive species remained significantly enhanced, even at sites of high RP, when forested reaches were present in upstream reaches. These probably provided a source of organisms for downstream recolonization and amelioration of effects at high RP.

Mercury resistance in bacterial strains isolated from tailing ponds in a gold mining area near El Callao (Bolívar State, Venezuela).

Bacterial resistance to mercury (Hg) was investigated in strains isolated from Hg-contaminated tailing ponds located in the gold mining area of El Callao (Bolívar State, Venezuela). High frequencies of resistance were detected to both inorganic-Hg and organomercurials among these strains. A broad range of resistance levels was observed when determining minimal inhibitory concentrations of Hg(2+). Some strains were able to grow in liquid medium containing 25 muM: Hg(2+), whereas others grew at 300 muM: Hg(2+). Of 190 Hg-resistant strains tested, 58.2% were additionally shown to be resistant to ampicillin (40 mg/L), 33.3% to chloramphenicol (30 mg/L), 24.9% to streptomycin (30 mg/L), 23.3% to tetracycline (30 mg/L), and 1.6% to kanamycin (30 mg/L). Furthermore, we found that 20% of the Hg-resistant strains were simultaneously resistant to as many as four of these antibiotics, at the concentrations tested. The presence of large plasmids in 62.9% of 53 Hg-resistant strains screened prompted us to investigate the horizontal transfer of resistance determinants. Mating experiments were performed using Escherichia coli and Pseudomonas aeruginosa as recipient strains. The results obtained confirmed that indigenous Hg-resistant bacteria colonizing the tailing ponds can effectively transfer the phenotype to potentially pathogenic species.

Recombinant environmental libraries provide access to microbial diversity for drug discovery from natural products.

To further explore possible avenues for accessing microbial biodiversity for drug discovery from natural products, we constructed and screened a 5,000-clone "shotgun" environmental DNA library by using an Escherichia coli-Streptomyces lividans shuttle cosmid vector and DNA inserts from microbes derived directly (without cultivation) from soil. The library was analyzed by several means to assess diversity, genetic content, and expression of heterologous genes in both expression hosts. We found that the phylogenetic content of the DNA library was extremely diverse, representing mostly microorganisms that have not been described previously. The library was screened by PCR for sequences similar to parts of type I polyketide synthase genes and tested for the expression of new molecules by screening of live colonies and cell extracts. The results revealed new polyketide synthase genes in at least eight clones. In addition, at least five additional clones were confirmed by high-pressure liquid chromatography analysis and/or biological activity to produce heterologous molecules. These data reinforce the idea that exploiting previously unknown or uncultivated microorganisms for the discovery of novel natural products has potential value and, most importantly, suggest a strategy for developing this technology into a realistic and effective drug discovery tool.