Parkin is a critical player in the effects of caffeine over mitochondrial quality control pathways during skeletal muscle regeneration in mice.

AIM: This study aimed to investigate the effects of caffeine on pathways associated with mitochondrial quality control and mitochondrial capacity during skeletal muscle regeneration, focusing on the role of Parkin, a key protein involved in mitophagy. METHODS: We used in vitro C2C12 myoblast during differentiation with and without caffeine in the medium, and we evaluated several markers of mitochondrial quality control pathways and myotube growth. In vivo experiments, we used C57BL/6J (WT) and Parkintm 1Shn lineage (Parkin-/- ) mice and injured tibial anterior muscle. The mice regenerated TA muscle for 3, 10, and 21 days with or without caffeine ingestion. TA muscle was used to analyze the protein content of several markers of mitochondrial quality pathways, muscle satellite cell differentiation, and protein synthesis. Furthermore, it analyzed mtDNA, mitochondrial respiration, and myofiber growth. RESULTS: C2C12 differentiation experiments showed that caffeine decreased Parkin content, potentially leading to increased DRP1 and PGC-1α content and altered mitochondrial population, thereby enhancing growth capacity. Using Parkin-/- mice, we found that caffeine intake during the regenerative process induces an increase in AMPKα phosphorylation and PGC-1α and TFAM content, changes that were partly Parkin-dependent. In addition, the absence of Parkin potentiates the ergogenic effect of caffeine by increasing mitochondrial capacity and myotube growth. Those effects are related to increased ATF4 content and activation of protein synthesis pathways, such as increased 4E-BP1 phosphorylation. CONCLUSION: These findings demonstrate that caffeine ingestion changes mitochondrial quality control during skeletal muscle regeneration, and Parkin is a central player in those mechanisms.

Position specific physical performance and running intensity fluctuations in elite women's football.

The purpose of the present study was to investigate the physical performance of elite female football players during match play along with transient alterations in running performance following 1- and 5-min univariate peak periods. 54 elite female players from four top-level Norwegian teams were monitored for one season (n = 393 match observations), and physical performance data collected using STATSport GPS APEX. Results revealed significant differences in physical performance between the positions during full match play, particularly between wide and central players. Both full backs (FBs) and wide midfielders (WMs) covered more total distance (TD), high-speed running distance (HSRD), and sprint distance (SpD) than center backs (CBs) (p < 0.05-0.001), while WMs also covered more HSRD than both central midfielders (CMs) (p < 0.01) and forwards (FWs) (p < 0.05), and more acceleration -and deceleration distance (Accdist and Decdist ) than both CBs and CMs (p < 0.01-0.001). A similar pattern was observed for the peak period analysis, with FBs and WMs covering more SpD in peak 1 min than CBs and CM (p < 0.001) and more SpD in peak 5-min than CBs, CMs, and FWs (p < 0.001). Irrespective of the variable analyzed, greater distances were covered during the peak 5-min period than in the next-5 and mean 5-min periods (p < 0.001). Significant (p < 0.001), but small to trivial (Cohen's Dz : 0.07-0.20), decreases in distance covered were also observed for each variable following each univariate peak 5-min period. In conclusion, practitioners should account for differences in physical performance when developing training programs for female football players and be aware of transient reductions in physical performance following univariate peak 1- and 5-min periods. Specifically, the very high intensity in 1-min peak periods adds support to the principal of executing speed endurance activities during training to mirror and be prepared for the physical demands of match play.

The E3 ligase MuRF2 plays a key role in the functional capacity of skeletal muscle fibroblasts.

Fibroblasts are a highly heterogeneous population of cells, being found in a large number of different tissues. These cells produce the extracellular matrix, which is essential to preserve structural integrity of connective tissues. Fibroblasts are frequently engaged in migration and remodeling, exerting traction forces in the extracellular matrix, which is crucial for matrix deposition and wound healing. In addition, previous studies performed on primary myoblasts suggest that the E3 ligase MuRF2 might function as a cytoskeleton adaptor. Here, we hypothesized that MuRF2 also plays a functional role in skeletal muscle fibroblasts. We found that skeletal muscle fibroblasts express MuRF2 and its siRNA knock-down promoted decreased fibroblast migration, cell border accumulation of polymerized actin, and down-regulation of the phospho-Akt expression. Our results indicated that MuRF2 was necessary to maintain the actin cytoskeleton functionality in skeletal muscle fibroblasts via Akt activity and exerted an important role in extracellular matrix remodeling in the skeletal muscle tissue.

The E3 ligase MuRF2 plays a key role in the functional capacity of skeletal muscle fibroblasts

Fibroblasts are a highly heterogeneous population of cells, being found in a large number of different tissues. These cells produce the extracellular matrix, which is essential to preserve structural integrity of connective tissues. Fibroblasts are frequently engaged in migration and remodeling, exerting traction forces in the extracellular matrix, which is crucial for matrix deposition and wound healing. In addition, previous studies performed on primary myoblasts suggest that the E3 ligase MuRF2 might function as a cytoskeleton adaptor. Here, we hypothesized that MuRF2 also plays a functional role in skeletal muscle fibroblasts. We found that skeletal muscle fibroblasts express MuRF2 and its siRNA knock-down promoted decreased fibroblast migration, cell border accumulation of polymerized actin, and down-regulation of the phospho-Akt expression. Our results indicated that MuRF2 was necessary to maintain the actin cytoskeleton functionality in skeletal muscle fibroblasts via Akt activity and exerted an important role in extracellular matrix remodeling in the skeletal muscle tissue.

Thyroid hormone upregulates MDM2 in rat type I fibre: Implications for skeletal muscle mass regulation.

AIM: Based upon a microarray assay, we have identified that triiodothyronine (T3) upregulates MDM2 gene expression in the rat skeletal muscle. As MDM2 protein is an E3 ligase, we hypothesized that this enzyme could play a role in T3 effects on skeletal muscle mass control. METHODS: To test our hypothesis, male rats (2 months old) were randomly assigned into the following groups: intact controls, treated with 20 physiological doses of T3 for 0.5, 1 and 7 days, or with 5, 20 and 50 physiological doses of T3 for 7 days. For in vitro experiments, myotubes and C2C12 cells were treated with T3 for 3 days. RESULTS: After validation of the microarray finding throughout RT-PCR and confirmation that T3 induces increases in MDM2 protein expression in a dose-dependent manner, we observed that MDM2 was upregulated by T3 exclusively in fibre type I. Moreover, detailed histological evaluation showed that MDM2 overexpression distributes punctiformily along the cross section of the fibre and also inside nuclei. MDM2 colocalizes with PAX7 in control muscle and T3 downregulates this myogenic factor. Pharmacological inhibition of MDM2 in cultured myotubes caused a severe decrease in their diameter (~35%, P < .001 vs Control), enhancing the effect of T3 (from ~12% to ~35%, P < .001) alone upon myotube diameter and mRNA levels of atrogenes. Finally, we observed that FOXO3 (MDM2 target) is kept outside the nucleus under T3 stimulation. CONCLUSION: Our results indicate that MDM2 might be involved in the pro-trophic effects of T3 in skeletal muscle.

Histological and profilometric evaluation of the root surface after instrumentation with a new piezoelectric device - ex vivo study.

OBJECTIVES: An ex vivo model was designed to profilometrically and histologically assess root changes resulting from scaling with a new ultrasonic device, designed for bone piezoelectric surgery, in comparison with curettes. METHODS: Three groups of 10 periodontal hopeless teeth were each subjected to different root instrumentation: Gracey curettes (CUR); ultrasonic piezoelectric device, Perio 100% setting, level 8 (P100); and ultrasonic piezoelectric device Surg 50% setting, level 1 (S50). After extraction, all teeth were photographed to visually assess the presence of dental calculus. The treated root surfaces were profilometrically evaluated (Ra, Rz, Rmax). Undecalcified histological sections were prepared to assess qualitative changes in cementum thickness. Statistical analysis was carried out using one-way anova test with a significance level of 95%. RESULTS: Both instruments proved to be effective in the complete removal of calculus. The CUR group presented the lowest Ra [2.28 µm (±0.58)] and S50 the highest [3.01 µm (±0.61)]. No statistically significant differences were detected among the three groups, for Ra, Rz and Rmax. Histologically, there was a cementum thickness reduction in all groups, being higher and more irregular in S50 group. CONCLUSIONS: Within the limits of this study, there were no statistically significant differences in roughness parameters analyzed between curettes and the ultrasonic piezoelectric unit. This new instrument removes a smaller amount of cementum, mainly at the Perio 100% power setting, which appears to be the least damaging. The ultrasonic device is effective in calculus removal, proving to be as effective as curettes.

Validation of a dental image-analyzer tool to measure the radiographic defect angle of the intrabony defect in periodontitis patients.

BACKGROUND AND OBJECTIVE: A report describing the software Dental Image Analyzer (DIA) was published in this journal in 2009. A new function - measurement of the periodontal intrabony defect angle - was added to the software in 2010. The purpose of this study was to investigate whether measurements of the radiographic intrabony defect angle using digital radiographs and the newly developed DIA tool were comparable with measurements obtained using the conventional protractor method. MATERIAL AND METHODS: The baseline radiographic defect angle of intrabony defects was measured conventionally, using a protractor, in 60 selected teeth from 47 patients and then digitally using the newly developed DIA tool. The measurements were made independently by four experienced dentists. The radiographic defect angle of intrabony defects was measured after the three anatomical landmarks were identified, namely the cemento-enamel junction, the top of the crest and the bottom of the defect. RESULTS: Both methods showed a high interexaminer reliability for measurements of the radiographic defect angle of intrabony defects (intraclass correlation coefficient > 0.97). Moreover, both methods showed high reliability (intraclass correlation coefficient > 0.96). On the other hand, the new DIA tool, compared with the conventional method, exhibited high sensitivity (0.92) and high specificity (0.91) in selecting defects of ≥ 37° or < 37°. Analysis of the time taken for measurements revealed significant differences between the two methods, with the protractor method being more time consuming. CONCLUSIONS: This study provided evidence for the lack of a significant difference between the conventional method and the DIA tool for radiographic measurement of intrabony defects. However, digital analysis was significantly faster.

Diversity in UV sensitivity and recovery potential among bacterioneuston and bacterioplankton isolates.

AIMS: To assess the variability in UV-B (280-320 nm) sensitivity of selected bacterial isolates from the surface microlayer and underlying water of the Ria de Aveiro (Portugal) estuary and their ability to recover from previous UV-induced stress. METHODS AND RESULTS: Bacterial suspensions were exposed to UV-B radiation (3·3 W m⁻²). Effects on culturability and activity were assessed from colony counts and (3) H-leucine incorporation rates, respectively. Among the tested isolates, wide variability in UV-B-induced inhibition of culturability (37·4-99·3%) and activity (36·0-98·0%) was observed. Incubation of UV-B-irradiated suspensions under reactivating regimes (UV-A, 3·65 W m⁻²; photosynthetic active radiation, 40 W m⁻²; dark) also revealed diversity in the extent of recovery from UV-B stress. Trends of enhanced resistance of culturability (up to 15·0%) and enhanced recovery in activity (up to 52·0%) were observed in bacterioneuston isolates. CONCLUSIONS: Bacterioneuston isolates were less sensitive and recovered more rapidly from UV-B stress than bacterioplankton isolates, showing enhanced reduction in their metabolism during the irradiation period and decreased culturability during the recovery process compared to bacterioplankton. SIGNIFICANCE AND IMPACT OF THE STUDY: UV exposure can affect the diversity and activity of microbial communities by selecting UV-resistant strains and alter their metabolic activity towards protective strategies.

IFNG +874 T>A single nucleotide polymorphism is associated with leprosy among Brazilians.

Leprosy is a chronic infectious disease caused by Mycobacterium leprae, a low virulence mycobacterium, and the outcome of disease is dependent on the host genetics for either susceptibility per se or severity. The IFNG gene codes for interferon-γ (IFN-γ), a cytokine that plays a key role in host defense against intracellular pathogens. Indeed, single nucleotide polymorphisms (SNPs) in IFNG have been evaluated in several genetic epidemiological studies, and the SNP +874T>A, the +874T allele, more specifically, has been associated with protection against infectious diseases, especially tuberculosis. Here, we evaluated the association of the IFNG locus with leprosy enrolling 2,125 Brazilian subjects. First, we conducted a case-control study with subjects recruited from the state of São Paulo, using the +874 T>A (rs2430561), +2109 A>G (rs1861494) and rs2069727 SNPs. Then, a second study including 1,370 individuals from Rio de Janeiro was conducted. Results of the case-control studies have shown a protective effect for +874T carriers (OR(adjusted) = 0.75; p = 0.005 for both studies combined), which was corroborated when these studies were compared with literature data. No association was found between the SNP +874T>A and the quantitative Mitsuda response. Nevertheless, the spontaneous IFN-γ release by peripheral blood mononuclear cells was higher among +874T carriers. The results shown here along with a previously reported meta-analysis of tuberculosis studies indicate that the SNP +874T>A plays a role in resistance to mycobacterial diseases.

Genetic, epidemiological and biological analysis of interleukin-10 promoter single-nucleotide polymorphisms suggests a definitive role for -819C/T in leprosy susceptibility.

Leprosy is a complex infectious disease influenced by genetic and environmental factors. The genetic contributing factors are considered heterogeneous and several genes have been consistently associated with susceptibility like PARK2, tumor necrosis factor (TNF), lymphotoxin-alpha (LTA) and vitamin-D receptor (VDR). Here, we combined a case-control study (374 patients and 380 controls), with meta-analysis (5 studies; 2702 individuals) and biological study to test the epidemiological and physiological relevance of the interleukin-10 (IL-10) genetic markers in leprosy. We observed that the -819T allele is associated with leprosy susceptibility either in the case-control or in the meta-analysis studies. Haplotypes combining promoter single-nucleotide polymorphisms also implicated a haplotype carrying the -819T allele in leprosy susceptibility (odds ratio (OR)=1.40; P=0.01). Finally, we tested IL-10 production in peripheral blood mononuclear cells stimulated with Mycobacterium leprae antigens and found that -819T carriers produced lower levels of IL-10 when compared with non-carriers. Taken together, these data suggest that low levels of IL-10 during the disease outcome can drive patients to a chronic and unprotective response that culminates with leprosy.

Radicicol improves regeneration of skeletal muscle previously damaged by crotoxin in mice.

This work investigates the influence of heat shock proteins (HSPs) on necrosis and subsequent skeletal muscle regeneration induced by crotoxin (CTX), the major component of Crotalus durissus terrificus venom. Mice were treated with radicicol, a HSP inductor, followed by an intramuscular injection of CTX into the gastrocnemius muscle. Treated groups were sacrificed 1, 10 and 21 days after CTX injection. Muscle histological sections were stained with toluidine blue and assayed for acid phosphatase or immunostained with either neuronal cell adhesion molecule (NCAM) or neonatal myosin heavy chain (MHCn). Muscle samples were also submitted to Western blotting analysis. The results show that CTX alone and CTX combined with radicicol induced a similar degree of myofiber necrosis. CTX-injured muscles treated with radicicol had increased cross-sectional areas at 10 and 21 days post-lesion compared with untreated CTX-injured muscles. Additionally, radicicol significantly increased the number of NCAM-positive satellite cells in the gastrocnemius at one day post-CTX injury. CTX-injured muscles treated with radicicol contained more MHCn-positive regenerating myofibers compared with untreated CTX-injured muscles. These results suggest that HSPs contribute to the regeneration of myofibers damaged by CTX. Additionally, further studies should investigate the potential therapeutic effects of radicicol in skeletal muscles affected by Crotalus venom.

Evidence of species succession during chlorobenzene biodegradation.

We have previously reported the disappearance of a specific strain degrading chlorobenzene from a functionally stable bioreactor. In the present work, we investigated this species succession and isolated a new dominant strain, identified as Pandoraea pnomenusa sp. strain MCB032. A specific 16S rRNA-targeted oligonucleotide probe was designed and validated to identify strain MCB032 using fluorescence in situ hybridisation (FISH). The results confirmed the presence of strain MCB032 in samples collected over time, and showed that it was primarily located within the biofilm. Denaturing gradient gel electrophoresis (DGGE) provided evidence that the species succession occurred early in the operating period. The application of these biomolecular tools highlighted the remarkable stability of this new strain during the 15 months of reactor operation. The succession was attributed to the competitive kinetic behaviour of strain MCB032, which exhibited faster growth (micro(max) = 0.34 h(-1)) and higher substrate affinity (K(s) = 0.35 mg L(-1)) than strain JS150. Finally, this study contributed to the characterisation of the recently established Pandoraea genus, an emerging group in the biodegradation field.

Effect of calcineurin inhibitors on myotoxic activity of crotoxin and Bothrops asper phospholipase A2 myotoxins in vivo and in vitro.

Previous studies have shown that calcineurin activity plays a critical role in the myotoxic activity induced by crotoxin (CTX), a group II phospholipase A(2) (PLA(2)) with neurotoxic and myotoxic actions. In order to address whether calcineurin is also important for the activity of non-neurotoxic group II PLA(2) myotoxins we have compared the effects of calcineurin inhibition on the myotoxic capacity of CTX and the non-neurotoxic PLA(2)s, myotoxin II (Mt II) and myotoxin III (Mt III) from Bothrops asper venom. Rats were treated with cyclosporin A (CsA) or FK506, calcineurin inhibitors, and received an intramuscular injection of either CTX, Mt II or Mt III into the tibialis anterior. Animals were killed 24 h after injection of toxins. Tibialis anterior was removed and stored in liquid nitrogen. Myofibers in culture were also treated with CsA or FK506 and exposed to CTX, Mt II and Mt III. It was observed that, in contrast to CTX, CsA and FK506 do not attenuate myotoxic effects induced by both Mt II and Mt III in vivo and in vitro. The results of the present study suggest that calcineurin is not essential for the myotoxic activity of Mt II and Mt III, indicating that distinct intracellular pathways might be involved in myonecrosis induced by neurotoxic CTX and non-neurotoxic Bothrops sp. PLA(2) myotoxins. Alternatively, calcineurin dependent fast fiber type shift might render the muscle resistant to the action of CTX, without affecting its susceptibility to Bothrops sp. myotoxins.

Strain stability in biological systems treating recalcitrant organic compounds.

The availability of molecular probing technology in recent years has facilitated investigation of microbial community composition during bio-treatment of organic wastes. Particularly, it has allowed the study of microbial culture stability and correlation between stability and treatment performance. However, most studies to date have only addressed mixed cultures and there is limited information regarding single strain stability. Here we have investigated the microbial community dynamics in two bioreactors, each inoculated with a pure bacterial strain capable of degrading a recalcitrant substrate, namely Xanthobacter aut. GJ10 degrading 1,2-dichloroethane (DCE) and Burkholderia sp. JS150 degrading monochlorobenzene (MCB). Universal and strain specific 16S rRNA oligonucleotide probes were designed and used to follow strain stability. The bioreactor fed with DCE was functionally stable and the percentage of GJ10 cells in the community remained high (around 95% of total cells) throughout, even after introduction of foreign microorganisms. The bioreactor fed with MCB was also functionally stable, but in contrast to the DCE bioreactor, probing results revealed the disappearance of strain JS150 from the bioreactor within a week. The difference in behavior between the two systems is attributed to the specific pathway required to degrade DCE.

Quarto de iodoterapia / Iodotherapy's room

Este trabalho tem por objetivo apresentar uma descrição dos procedimentos necessários para montagem, instalação e manutenção de um quarto de iodoterapia, tendo como meta principal uma melhor relação entre os fatores: necessidades do paciente e adequação com as normas de proteção radiológica estabelecidas pela Comissão Nacional de Energia Nuclear.

This articles has the objective to show the description of rules necessaries to assemble, to installation and mainterence the iodotherapy's room; and having the principal aim, get a better relacion between the factors: necessities of the pacient and the adaptation with the norms of protection radiologic established by the National Comission Nuclear Energy.

[Transverse myelitis]. / Mielite transversa.

The authors report three clinical cases of acute transverse myelitis in young patients with emphasis given to the seriousness of this kind of pathology, the need to exclude a potentially treatable cause and the controversial corticosteroid treatment.

[Systemic lupus erythematosus in the elderly]. / Lupus eritematoso sistémico no idoso.

The authors report two clinical cases of systemic lupus erythematosus, both in patients over 65 years of age. The discussion includes a brief overview of recent publications on this condition in geriatric patients. Some aspects of systemic lupus in elderly persons are suggested and commented on.

[Adenocarcinoma of the colon disclosed by endocarditis caused by Streptococcus bovis]. / Adenocarcinoma do colon revelado por endocardite por estreptococos bovis.

The association of Streptococcus bovis endocarditis with colonic neoplasms has been well documented. We describe a patient in whom the finding of a Str. bovis endocarditis stimulated investigation which resulted in the diagnosis and surgical treatment of an adenocarcinoma of the colon.