Endurance exercise associated with a fructooligosaccharide diet modulates gut microbiota and increases colon absorptive area.

BACKGROUND AND AIM: Fructooligosaccharide (FOS) supplementation can stimulate beneficial intestinal bacteria growth, but little is known about its influence on training performance. Therefore, this study analyzed FOS and exercise effects on gut microbiota and intestinal morphology of C57Bl/6 mice. METHODS: Forty male mice were divided into four groups: standard diet-sedentary (SDS), standard diet-exercised (SDE), FOS supplemented (7.5% FOS)-sedentary (FDS), and FOS supplemented-exercised (FDE), n = 10 each group. Exercise training consisted of 60 min/day, 3 days/week, for 12 weeks. RESULTS: SDE and FDE groups had an increase in aerobic performance compared to the pretraining period and SDS and FDS groups (P < 0.01), respectively. Groups with FOS increased colonic crypts size (P < 0.05). The FDE group presented rich microbiota (α-diversity) compared to other groups. The FDE group also acquired a greater microbial abundance (ß-diversity) than other groups. The FDE group had a decrease in the Ruminococcaceae (P < 0.002) and an increase in Roseburia (P < 0.003), Enterorhabdus (P < 0.004) and Anaerotruncus (P < 0.006). CONCLUSIONS: These findings suggest that aerobic exercise associated with FOS supplementation modulates gut microbiota and can increase colonic crypt size without improving endurance exercise performance.

Effects of 12†weeks of resistance training on rat gut microbiota composition.

In addition to its health benefits, exercise training has been noted as a modulator of the gut microbiota. However, the effects of resistance training (RT) on gut microbiota composition remain unknown. Wistar rats underwent 12 weeks of RT. Body mass, glucose tolerance, visceral body fat, triglyceride concentration and food consumption were evaluated. The gut microbiota was analyzed by 16S rRNA gene sequencing. Rats that underwent RT showed lower body mass (P=0.0005), lower fat content (P=0.02) and better glucose kinetics (P=0.047) when compared with the control. Improvements in the diversity and composition of the gut microbiota were identified in the RT group. The relative abundance of Pseudomonas, Serratia and Comamonas decreased significantly after 12 weeks of RT (P<0.001). These results suggest that RT has the potential to enhance the diversity of the gut microbiota and improve its biological functions.

Changes in Compliance With Physical Activity Guidelines and Cardiovascular Disease Mortality.

BACKGROUND: The effects of compliance with the US Physical Activity (PA) Guidelines and changes in compliance over time on cardiovascular disease (CVD) mortality are unknown. METHODS: Male participants in the Aerobics Center Longitudinal Study (n = 15,411; 18-100 y) reported leisure-time PA between 1970 and 2002. The frequency of and time spent in PA were converted into metabolic equivalent minutes per week. The participants were classified into remained inactive, became active, became inactive, or remained active groups according to their achievement of the PA guidelines along the follow-up, equivalent here to at least 500 metabolic equivalent minutes of PA per week. Cox regression adjusted for different models was used for the analyses, using age, body mass index, smoking and drinking status, hypertension, diabetes, hypercholesterolemia, and parental history of CVD. RESULTS: Over a mean follow-up of 6.2 years, 439 CVD deaths occurred. Consistently meeting the PA guidelines, compared with not meeting, was associated with a 54% (95% confidence interval, 0.32-0.67) decreased risk of CVD mortality. After controlling for all potential confounders, the risk reduction was 47% (95% confidence interval, 0.36-0.77). CONCLUSIONS: Maintaining adherence to the PA guidelines produces substantial reductions in the risk of CVD deaths in men. Furthermore, discontinuing compliance with the guidelines may offset the beneficial effects on longevity.

The use of host defense peptides in root canal therapy in rats.

OBJECTIVES: In order to evaluate host defense peptides (HDPs) HHC-10 and synoeca-MP activity in in vitro osteoclastogenesis process and in vivo induced apical periodontitis, testing the effect of molecules in the inflammatory response and in apical periodontitis size/volume after root canal treatment. MATERIALS AND METHODS: In vitro osteoclastogenesis was assessed on bone marrow cell cultures extracted from mice, while in vivo endodontic treatment involved rats treated with Ca(OH)2 or HDPs. In vitro osteoclasts were subjected to TRAP staining, and in vivo samples were evaluated by radiographic and tomographic exams, as well as histologic analysis. RESULTS: None of the substances downregulated the in vitro osteoclastogenesis. Nevertheless, all treatments affected the average of apical periodontitis size in rats, although only teeth treated with HDPs demonstrated lower levels of the inflammatory process. These results demonstrated the in vivo potential of HDPs. Radiographic analysis suggested that HHC-10 and synoeca-MP-treated animals presented a similar lesion size than Ca(OH)2-treated animals after 7-day of endodontic treatment. However, tomography analysis demonstrated smaller lesion volume in synoeca-MP-treated animals than HHC-10 and Ca(OH)2-treated animals, after 7 days. CONCLUSIONS: These molecules demonstrated an auxiliary effect in endodontic treatment that might be related to its immunomodulatory ability, broad-spectrum antimicrobial activity, and possible induction of tissue repair at low concentrations. These results can encourage further investigations on the specific mechanisms of action in animal models to clarify the commercial applicability of these biomolecules for endodontic treatment. CLINICAL SIGNIFICANCE: HDPs have the potential to be adjuvant substances in endodontic therapy due to its potential to reduce inflammation in apical periodontitis.

Host defense peptide IDR-1002 associated with ciprofloxacin as a new antimicrobial and immunomodulatory strategy for dental pulp revascularization therapy.

Regenerative therapies such as dental pulpal revascularization appear as an option for traumatized immature permanent teeth. However, the triple antibiotic paste - TAP (metronidazole, minocycline, and ciprofloxacin), used for these therapies, can generate cytotoxicity and dentin discoloration. In contrast, host defense peptides (HDPs) are promising antimicrobial and immunomodulatory biomolecules for dentistry. This study aimed to evaluate in vitro the antimicrobial activity (against Staphylococcus aureus and Enterococcus faecalis) and the immunomodulatory potential (by the evaluation of IL-1α, IL-6, IL-12, IL-10, TNF-α and NO, in RAW 264.7 macrophages and IL-6, TGF-ß and NO, in L929 fibroblast) of synthetic peptides (DJK-6, IDR-1018, and IDR-1002), compared to TAP in an in vitro infection model containing heat-killed antigens from E. faecalis and S. aureus. Furthermore, the synergistic potential of ciprofloxacin and IDR-1002 was evaluated by checkerboard. Ciprofloxacin was the best antimicrobial of TAP, besides acting in synergism with IDR-1002. TAP was pro-inflammatory (p < 0.05), while the association of ciprofloxacin and IDR-1002 presented an anti-inflammatory profile mainly in the presence of both heat-killed antigens (p < 0.05). Based on these results, ciprofloxacin associated with IDR-1002 may demonstrate an efficient antimicrobial and immunomodulatory action in this in vitro model. Further in vivo studies may determine the real potential of this combination.

High-Protein Diet Associated with Bocaiuva Supplementation Decreases Body Fat and Improves Glucose Tolerance in Resistance-Trained Rats.

High-protein diets (HPDs) are widely used for health and performance. However, the combination of whey protein and natural foods (i.e., fruits) is still unclear. Thus, we evaluated the role of supplemental HPD with Bocaiuva (Acrocomia sp.) in metabolic and body composition parameters of rats submitted to resistance training (RT). Wistar rats (203.3 ± 30 g) were randomly allocated to five groups: normoproteic control (CON, n = 5), sedentary high-protein (SH, n = 5), RT + H (trained high-protein [TH], n = 5), sedentary+Bocaiuva (SH+B, n = 4), and RT+Bocaiuva (TH+B, n = 4) diet groups. After 12 weeks of RT, the maximal strength increased in both trained groups (P < .05). The TH + B group had lower values of adiposity index (AI) (3.8 ± 0.7% vs. 6.8 ± 1.3%) and visceral fat (0.038 ± 0.004 g/g vs. 0.067 ± 0.012 g/g) compared with the SH group, respectively (P < .05). The other groups did not show differences in values of AI (CON, 5.4 ± 1.6%, TH, 5.4 ± 1.3%, and SH+B, 5.5 ± 1.2%). In addition, the fasting glucose of trained groups (TH, 106.0 ± 4.5, and TH+B, 100.4 ± 13.5 dL/mg) was significantly lower when compared with controls (SH, 120.0 ± 14.4, and SH+B, 119 ± 6.4 dL/mg) (P < .05). Bocaiuva combined with an HPD reduced visceral fat and AI in addition to improving glucose tolerance of rats submitted to RT.

Identification of the force-velocity curve on dynamic resistance exercise for rats.

The aim of this study was to identify force-velocity and power-velocity curves in climbing activity protocols, used as dynamic resistance exercise in rats. Eighteen 45-day-old male Wistar rats (weight = 211.9 ± 5.2 g) were evaluated. After familiarization to the climbing procedure, the animals performed an incremental climbing test (load relative to 75% of the body mass at first stage, followed by 30 g increments with and 120 s recovery between climbs) to determine the maximum carrying capacity (MCC). After this, the animals climbed with different loads (without load, 10%, 20%, 30%, 40%, 50%, 75%, 90%, and 100% of MCC) with 120 s recovery between climbs. Time for each climb was recorded to calculate the mechanical power. The peak power was reached at 30% of MCC. For the force-velocity curve, an inversely proportional relation was observed between force and velocity, as expected, greater forces were expressed in lower velocities. Therefore, our results suggest that training at 30% of MCC should be encouraged aiming the target for greater power output and 90%-100% of MCC should be the load aiming for strength training in climbing activities for rats.

Synergistic activity of chlorhexidine and synoeca-MP peptide against Pseudomonas aeruginosa.

‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬This study aims to evaluate the in vitro antimicrobial and immunomodulatory activities and cytotoxicity of chlorhexidine (CHX) and synoeca-MP peptide alone or in combination against Pseudomonas aeruginosa. The antimicrobial property was evaluated by the determination of minimal inhibitory concentration, minimum bactericidal concentration, and planktonic bacteria and biofilm inhibition. Immunomodulatory activity was determined by enzyme-linked immunosorbent assay and nitric oxide production by the Griess reaction method. According to the results, synoeca-MP combined with CHX demonstrated antimicrobial effectiveness compared with its isolated use, in addition to immunomodulatory activity (upregulating MPC-1 and tumor necrosis factor-α and downregulating nitric oxide and interleukin-10). In this context, it is expected that the substances, together, could be capable of controlling bacterial infection and dissemination, besides potentiating macrophages' immune response against the studied microorganism. Moreover, reducing the CHX concentration by the addition of synoeca-MP peptide may, in a beneficial way, minimize the undesirable effects of both, CHX and synoeca-MP in a clinical setting.

Limited Effects of Low-to-Moderate Aerobic Exercise on the Gut Microbiota of Mice Subjected to a High-Fat Diet.

Several studies have indicated that diet and exercise may modulate the gut microbiota in obese subjects. Both interventions were shown to alter the microbiota orthogonally. However, this relationship has not been fully explored. This study analyzed the effects of low-to-moderate aerobic training on the fecal microbiota of mice subjected to a high-fat diet (HFD). Here, 40 male mice (C57Bl/6) were divided into two groups with standard diet (SD; 12.4% lipid) and HFD (60.3% lipid) for four months. These groups were divided into four, named SD control, HF control, SD trained and HF trained. All animals were submitted to an incremental test to estimate low-to-moderate maximum speed. Training consisted of 30 min·day-1, 5 days/week, for 8 weeks. The HFD increased the body weight (p < 0.0001) and adiposity index (p < 0.05). HFD also negatively influenced performance in exercise training. Moreover, the diversity of gut microbiota was reduced by the HFD in all groups. A low-to-moderate exercise was ineffective in modulating the gut microbiota composition in mice subjected to HFD. These findings suggest that two months of low-to-moderate exercise does not achieve a preponderant modulatory effect on shaping microbiota when submitted to the high-fat diet.

Antimicrobial and proinflammatory effects of two vipericidins.

Hospital infections allied to bacterial resistance to antibiotics have become a major worldwide problem. In this context, antimicrobial peptides (AMPs) are presented as an alternative in the control of these resistant organisms. Besides antimicrobial effects, these molecules play a crucial role in immunity by acting as immunomodulators. These peptides can activate inflammatory cells to produce pro- and anti-inflammatory mediators. In this study we will show the activity against multi-drug resistant bacteria (MDRB) of two cathelicidins from South American pit vipers Bothrops atrox and Crotalus durissus terrificus, named batroxicidin and crotalicidin. It was observed that both peptides showed activity against MDRB and presented no hemolytic or cytotoxic activity. In addition, the ability of peptides to modulate the production of cytokines TNF-α, IL-10 and IL-6 was analyzed using Raw 264.7 cells in the presence of IFN-γ stimuli, and multi-resistant E. coli and K. pneumoniae antigens. An up-expression or down-expression of TNF-α, as well as the IL-10 mediator, was observed. The cytokine IL-6, on the other hand, presented only a down-regulation for Raw 264.7 cell groups. In conclusion, the results demonstrate that both peptides presented a predominantly proinflammatory characteristic to the inflammatory mediators dosed. Overall, even presenting a proinflammatory characteristic, these peptides are still promising for future research and development of new potential antimicrobial molecules.

Corrigendum: Two Consecutive Days of Extreme Conditioning Program Training Affects Pro and Anti-inflammatory Cytokines and Osteoprotegerin without Impairments in Muscle Power.

[This corrects the article DOI: 10.3389/fphys.2016.00260.].

Interplay between circulating nitric oxide and interleukin-17 in elderly outpatients with non-inflammatory conditions.

Nitric oxide (NOx) availability in biological systems is associated with either favorable or unfavorable outcomes. In this sense, several studies bring about evidence that unbalanced NOx production may be underlying to the pathophysiology of vascular disorders. Our study investigated the possible association of clinical, biochemical and inflammatory variables with total circulating levels of NOx in elderly patients devoid of major inflammatory conditions. Clinical (demographics, lifestyle, anthropometry, pressoric traits) and biochemical characteristics (lipemic, glycemic and hormonal profiles) were assessed from 168 geriatrics outpatients eligible for primary care for age-related disorders. Furthermore, circulating levels of 10 inflammatory mediators and of NOx were measured. Correlation tests analyzed categorical or continuous traits according to serum NOx and found no association between NOx and any of the clinical or laboratory data but a negative correlation between plasma NOx concentrations and levels of the immune mediator IL17a (r = -0.236; P = 0.004). Evidence for a correlation between circulating NOx and IL17 is already present in the literature, mostly from studies conducted under inflammatory conditions. Our hypothesis is that such negative correlation can be attributed to an endogenous homeostatic system that IL17 production by the constitutively produced NOx from the vascular endothelium.

Effects of Acute Aerobic Exercise on Rats Serum Extracellular Vesicles Diameter, Concentration and Small RNAs Content.

Physical exercise stimulates organs, mainly the skeletal muscle, to release a broad range of molecules, recently dubbed exerkines. Among them, RNAs, such as miRNAs, piRNAs, and tRNAs loaded in extracellular vesicles (EVs) have the potential to play a significant role in the way muscle and other organs communicate to translate exercise into health. Low, moderate and high intensity treadmill protocols were applied to rat groups, aiming to investigate the impact of exercise on serum EVs and their associated small RNA molecules. Transmission electron microscopy, resistive pulse sensing, and western blotting were used to investigate EVs morphology, size distribution, concentration and EVs marker proteins. Small RNA libraries from EVs RNA were sequenced. Exercise did not change EVs size, while increased EVs concentration. Twelve miRNAs were found differentially expressed after exercise: rno-miR-128-3p, 103-3p, 330-5p, 148a-3p, 191a-5p, 10b-5p, 93-5p, 25-3p, 142-5p, 3068-3p, 142-3p, and 410-3p. No piRNA was found differentially expressed, and one tRNA, trna8336, was found down-regulated after exercise. The differentially expressed miRNAs were predicted to target genes involved in the MAPK pathway. A single bout of exercise impacts EVs and their small RNA load, reinforcing the need for a more detailed investigation into EVs and their load as mediators of health-promoting exercise.

The effects of glucose concentrations associated with lipopolysaccharide and interferon-gamma stimulus on mediators' production of RAW 264.7 cells.

Diabetes mellitus (DM) is a metabolic disorder that results in the impairment of the metabolism of carbohydrates, proteins and lipids. It can give rise to various complications, mainly caused by chronic exposure of cells to high glucose concentrations, including changes in the immune response processes. The aim of this study was to verify the chemokine and cytokines production profile in the presence of different glucose concentrations and infection/inflammatory stimuli. To this end, cell viability and the production of chemokines, cytokines and nitric oxide (NO) were analyzed in RAW 264.7 cell culture. Results demonstrated that there was no change in cell viability after 6, 24 and 72 h. Different stimuli were unable to modify the monocyte chemoattractant protein (MCP)-1 and tumor necrosis factor (TNF)-α production. Groups stimulated with lipopolysaccharides (LPS) and LPS and recombinant interferon (rIFN)-γ down-regulated interleukin (IL)-1α, IL-10 and IL-12 and up-regulated IL-6 production. NO production maintained a pattern of increase, according to the increase in glucose concentrations, reaching its peak at 72 h. In summary, the results demonstrated that high glucose concentrations alone may be sufficient to alter the in vitro mediators' production of RAW 264.7 cells.

Antimicrobial and immunomodulatory activity of host defense peptides, clavanins and LL-37, in vitro: An endodontic perspective.

Endodontic treatment is mainly based on root canal disinfection and its failure may be motivated by microbial resistance. Endodontic therapy can be benefitted by host defense peptides (HDPs), which are multifunctional molecules that act against persistent infection and inflammation. This study aimed to evaluate the antimicrobial, cytotoxic and immunomodulatory activity of several HDPs, namely clavanin A, clavanin A modified (MO) and LL-37, compared to intracanal medication Ca(OH)2. HDPs and Ca(OH)2 were evaluated by: (1) antimicrobial assays against Candida albicans and Enterococcus faecalis, (2) cytotoxicity assays and (3) cytokine tumor necrosis factor (TNF)-α, monocyte chemoattractant protein (MCP)-1, interleukin (IL)-1α, IL-6, IL-10 and IL-12 and nitric oxide (NO) production by RAW 264.7 cells incubated with or without heat-killed (HK) C. albicans or E. faecalis combined or not with interferon-γ. The minimum inhibitory concentration (MIC) was established only for E. faecalis (LL-37, 57µM). Considering cytotoxicity, clavanin MO was able to reduce cell viability in many groups and demonstrated lowest LC50. The Ca(OH)2 up-regulated the production of MCP-1, TNF-α, IL-12 and IL-6 and down-regulated IL-1α, IL-10 and NO. Clavanins up-regulated the TNF-α and NO and down-regulated IL-10 production. LL-37 demonstrated up-regulation of IL-6 and TNF-α production and down-regulation in IL-10 and NO production. In conclusion, LL-37 demonstrated better antibacterial potential. In addition, Ca(OH)2 demonstrated a proinflammatory response, while the HDPs modulated the inflammatory response from non-interference with the active cytokines in the osteoclastogenesis process, probably promoting the health of periradicular tissues.

Fractionated Concurrent Exercise throughout the Day Does Not Promote Acute Blood Pressure Benefits in Hypertensive Middle-aged Women.

Hypertension is a chronic disease that affects about 30% of the world's population, and the physical exercise plays an important role on its non-pharmacological treatment. Anywise, the dose-response of physical exercise fractionation throughout the day demands more investigation, allowing new exercise prescription possibilities. Therefore, this study aimed to analyze the acute blood pressure (BP) kinetics after 1 h of exercises and the BP reactivity after different concurrent exercise (CE) sessions and its fractioning of hypertensive middle-aged women. In this way, 11 hypertensive women voluntarily underwent three experimental sessions and one control day [control session (CS)]. In the morning session (MS) and night session (NS), the exercise was fully realized in the morning and evening, respectively. For the fractionized session (FS), 50% of the volume was applied in the morning and the remaining 50% during the evening. The MS provided the greatest moments (p ≤ 0.05) of post-exercise hypotension (PEH) for systolic BP (SBP) and highest reduction of BP reactivity for SBP (~44%) and diastolic BP (DBP) (~59%) compared to CS (p ≤ 0.05). The findings of the present study have shown that MS is effective for PEH to SBP, as well as it promotes high quality of attenuation for BP reactivity, greater than the other sessions.

The Effects of Acute and Chronic Exercise on Skeletal Muscle Proteome.

Skeletal muscle plasticity and its adaptation to exercise is a topic that is widely discussed and investigated due to its primary role in the field of exercise performance and health promotion. Repetitive muscle contraction through exercise stimuli leads to improved cardiovascular output and the regulation of endothelial dysfunction and metabolic disorders such as insulin resistance and obesity. Considerable improvements in proteomic tools and data analysis have broth some new perspectives in the study of the molecular mechanisms underlying skeletal muscle adaptation in response to physical activity. In this sense, this review updates the main relevant studies concerning muscle proteome adaptation to acute and chronic exercise, from aerobic to resistance training, as well as the proteomic profile of natural inbred high running capacity animal models. Also, some promising prospects in the muscle secretome field are presented, in order to better understand the role of physical activity in the release of extracellular microvesicles and myokines activity. Thus, the present review aims to update the fast-growing exercise-proteomic scenario, leading to some new perspectives about the molecular events under skeletal muscle plasticity in response to physical activity. J. Cell. Physiol. 232: 257-269, 2017. © 2016 Wiley Periodicals, Inc.

Oxidative stability of sesame and flaxseed oils and their effects on morphometric and biochemical parameters in an animal model.

BACKGROUND: Sesame and flaxseed oils, which are rich in essential n-6 and n-3 polyunsaturated fatty acids, are widely consumed. We have determined the optical behavior with respect to the quality and identity of cold-pressed sesame and flaxseed oils. The effects of these oils and their combinations on metabolic parameters in animal models were also measured. RESULTS: Flaxseed oil emitted carotenoid fluorescence (500-650 nm), although it was more unstable than sesame oil, which had a larger induction period by the Rancimat method. The greater stability of sesame may be a result of the lower quantity of linolenic fatty acids. These oils were added to the feed of 56 rats, whereas animal fat was used for the control group. The sesame oil, flaxseed oil and sesame + flaxseed oils groups showed a significantly reduced adiposity index and blood glucose compared to the control group, whereas total cholesterol, high-density lipoprotein and triglycerides were lower in flaxseed oil and sesame + flaxseed oils (P < 0.05). Sesame + flaxseed oils had reduced levels of low-density lipoprotein and non-high-density lipoprotein (P < 0.05), indicating an anti-atherogenic effect in this group. CONCLUSION: Sesame oil was more stable than flaxseed oil. In an animal model, the diets with polyunsaturated fat sources proportions of 1:1 n-6:n-3 polyunsaturated fatty acids, improved the metabolic parameters, implying cardioprotective effects. © 2016 Society of Chemical Industry.

Two Consecutive Days of Crossfit Training Affects Pro and Anti-inflammatory Cytokines and Osteoprotegerin without Impairments in Muscle Power.

The aim of this study was to investigate the effects of two consecutive Crossfit® training sessions (24 h apart) designed to enhance work-capacity that involved both cardiovascular and muscular exercises on cytokines, muscle power, blood lactate and glucose. Nine male members of the CrossFit® community (age 26.7 ± 6.6 years; body mass 78.8 ± 13.2 kg; body fat 13.5 ± 6.2%; training experience 2.5 ± 1.2 years) completed two experimental protocols (24 h apart): (1) strength and power exercises, (2) gymnastic movements, and (3) metabolic conditioning as follows: 10 min of as many rounds as possible (AMRAP) of 30 double-unders and 15 power snatches (34 kg). The same sequence as repeated on session 2 with the following metabolic conditioning: 12 min AMRAP of: row 250 m and 25 target burpees. Serum interleukin-6 (IL-6), IL-10, and osteoprotegerin were measured before, immediately post and 24 h after workout of the day (WOD) 1, immediately post, 24 and 48 h after WOD 2. Peak and mean power were obtained for each repetition (back squat with 50% of 1 repetition maximum) using a linear position transducer measured before, immediately post and 24 h after WOD 1, immediately post and 24 h after WOD 2. Blood lactate and glucose were measured pre and immediately post WOD 1 and 2. Although both sessions of exercise elicited an significant increase in blood lactate (1.20 ± 0.41 to 11.84 ± 1.34 vs. 0.94 ± 0.34 to 9.05 ± 2.56 mmol/l) and glucose concentration (81.59 ± 10.27 to 114.99 ± 12.52 vs. 69.47 ± 6.97 to 89.95 ± 19.26 mg/dL), WOD 1 induced a significantly greater increase than WOD 2 (p ≤ 0.05). The training sessions elicited significant changes (p ≤ 0.05) in IL-6, IL-10 and osteoprotegerin concentration over time. IL-6 displayed an increase immediately after training WOD 1 [197 ± 109%] (p = 0.009) and 2 [99 ± 58%] (p = 0.045). IL-10 displayed an increase immediately after only WOD 1 [44 ± 52%] (p = 0.046), and decreased 24 and 48 h following WOD 2 (~40%; p = 0.018) as compared to pre-exercise values. Osteoprotegerin displayed a decrease 48 h following WOD 2 (~25%; p = 0.018) as compared with pre intervention. In conclusion, two consecutive Crossfit® training sessions increase pro/anti-inflammatory cytokines with no interference on muscle performance in the recovery period.

NanoUPLC-MS(E) proteomic analysis of osteoclastogenesis downregulation by IL-4.

Bone resorption is an important factor in bone homeostasis and imbalance can cause several diseases. In osteoimmunology, IL-4 has been described as an important factor in promoting M2 macrophage profile. In order to shed some light on the effect of IL-4 on osteoclast precursors in the presence of RANKL, cytokines and nitric oxide (NO) production and the proteomic profile were analyzed. The presence of IL-4 in in vitro osteoclastogenesis provides production of TNF-α, IL-1α, IL-1ß, IL-10 and IL-12 at basal cell levels. Regarding NO production, IL-4 was sufficient to increase the basal NO levels. Proteomic analyses identified 877 global proteins. IL-4 in in vitro RANKL-mediated osteoclastogenesis leads to the expression of 118 proteins. The presence of rIL-4 in in vitro rRANKL-mediated-osteoclastogenesis downregulated this process. However, the proteomics findings in the osteoclastogenesis demonstrated a much greater effect on osteoclast precursor cells than on RANKL-differentiated osteoclasts. These results suggest that the main effect of IL-4 in pre-osteoclast cells leads to a M2 macrophage activation, and this probably contributed to a reduction in osteoclastogenesis when both stimuli were used. This study noticed that IL-4 plays an important regulatory role in bone homeostasis due to its suppressive potential of precursor osteoclast cells.