Anti‐contractile effects of perivascular adipose tissue in thoracic aorta from rats fed a high‐fat diet: role of aerobic exercise training

The aim of the present study was to evaluate the effects of aerobic exercise training on perivascular adipose tissue (PVAT) function in thoracic aorta from rats fed a high‐fat diet. Aortic vascular reactivity was performed in sedentary (SD), trained (TR), sedentary high‐fat diet (SD‐HF), and trained high‐fat diet (TR‐HF) male Wistar rats in the absence (PVAT?) or in the presence (PVAT+) of thoracic PVAT. We also measured circulatory concentrations of leptin and tumour necrosis factor alpha (TNF‐α), as well as the protein expressions of TNF‐α receptor 1 (TNFR1) and inducible nitric oxide synthase (iNOS) on PVAT. In the SD‐HF group, the body weight, epididymal fat pad, thoracic PVAT, circulatory triglycerides, insulin, leptin and TNF‐α were increased when compared with the SD group, whereas exercise training reduced these values in TR‐HF group. The relaxing response curves to acetylcholine and sodium nitroprusside were not modified by either intervention (high‐fat diet or exercise training) or the presence of PVAT. The presence of PVAT had an anti‐contractile effect in response to serotonin in all groups. In SD‐HF group, the increased magnitude of anti‐contractile effects was in parallel with an up‐regulation of iNOS protein expression in PVAT without alteration in TNFR1. Exercise training was effective in normalizing the vascular reactivity in rings PVAT+ and in reducing the iNOS protein expression. Exercise training prevented the PVAT–induced alteration in thoracic aorta from rats fed a high‐fat diet.     

Identification of a novel mutation in the PAX9 gene in a family affected by oligodontia and other dental anomalies

The objective of the present work was to study the phenotype and the genotype of three generations of a family affected by oligodontia and other dental anomalies. These family members also presented systemic conditions such as hypercholesterolemia, hypothyroidism, diabetes mellitus, scoliosis, and congenital cardiovascular anomalies. Clinical evaluation, panoramic radiographs, and anamnestic data were used for dental analysis. DNA extraction was carried out from gum samples or buccal swabs. A mutation was identified in six subjects across three generations affected by oligodontia, as well as different phenotypical manifestations, both systemic and oral. The previously undescribed PAX9 mutation was observed in the paired box (exon 2); this was a heterozygote transition of C175 to T, implying the change of arginine 59 for a termination codon. These results strongly suggested that the identified mutation was the etiological cause of the oligodontia. However, in two family members affected by both hypodontia and peg-shaped upper lateral incisors, no mutations in the PAX9 and MSX1 genes were identified. This fact underscores the importance that other presently unknown genes and developmental factors have in tooth development and in the etiology of dental anomalies.     

Investigation of functional gene polymorphisms IL-1beta, IL-6, IL-10 and TNF-alpha in individuals with recurrent aphthous stomatitis

Recurrent aphthous stomatitis (RAS) is characterized by recurrent episodes of oral ulceration in an otherwise healthy individual. Some reports in the literature indicate that RAS may have immunological, psychological, genetic and microbiological bases. OBJECTIVE: The purpose of the present study was to investigate, using binary logistic regression analyses, a possible association between the functional IL-1beta +3954 (C/T), IL-6 -174 (G/C), IL-10 -1082 (G/A) and TNF-alpha -308 (G/A) genetic polymorphism and RAS in a sample of Brazilian patients, using a multivariate statistical analysis. DESIGN: Sixty-four consecutive subjects affected by minor and major forms of RAS and 64 healthy volunteers were genotyped. To investigate the association between the single nucleotide polymorphisms and risk of RAS, binary logistic regression models were fitted. The associations were expressed by odd ratios (ORs) and adjusted for age and gender, with the corresponding 95% CIs. P-values less than 0.05 were considered significant. RESULTS: A significant increase in the IL-1beta and TNF-alpha heterozygous genotypes were associated with an increased risk of RAS development (OR 2.40 and 3.07, respectively), in the multivariate model. CONCLUSION: Our findings demonstrate that polymorphisms of high IL-1beta and TNF-alpha production were associated with an increased risk of RAS development. Our findings also give additional support to a genetic basis for RAS pathogenesis.     

Confluent and reticulate papillomatosis of Gougerot-Carteaud and obesity: dermoscopic findings

Confluent and reticulated papillomatosis of Gougerot and Carteaud is a dermatosis that despite showing characteristic clinical signs is often poorly recognized and diagnosed. The authors present a case with extensive skin involvement, discuss its association with obesity and describe dermoscopic findings making the histopathological correlation.     

Discovering new in silico tools for antimicrobial peptide prediction

Antimicrobial peptides (AMPs) are important effectors of the innate immune system and play a vital role in the prevention of infections. Due to the increased emergence of new antibiotic-resistant bacteria, new drugs are constantly under investigation. AMPs in particular are recognized as promising candidates because of their modularity and wide antimicrobial spectrum. However, the mechanisms of action of AMPs, as well as their structure-activity relationships, are not completely understood. AMPs display no conserved three-dimensional structure and poor sequence conservation, which hinders rational design. Several bioinformatics tools have been developed to generate new templates with appealing antimicrobial properties with the aim of finding highly active peptide compounds with low cytotoxicity. The current tools reviewed here allow for the prediction and design of new active peptides with reasonable accuracy. However, a reliable method to assess the antimicrobial activity of AMPs has not yet been developed. The standardization of procedures to experimentally evaluate the antimicrobial activity of AMPs, together with the constant growth of current well-established databases, may allow for the future development of new bioinformatics tools to accurately predict antimicrobial activity.     

Arterial stiffness is increased in patients with type 1 diabetes without cardiovascular disease: a potential role of low-grade inflammation

OBJECTIVE

To investigate the relationship between arterial stiffness and low-grade inflammation in subjects with type 1 diabetes without clinical cardiovascular disease.

RESEARCH DESIGN AND METHODS

Sixty-eight patients with type 1 diabetes and 68 age- and sex-matched healthy subjects were evaluated. Arterial stiffness was assessed by aortic pulse wave velocity (aPWV). Serum concentrations of high-sensitivity C-reactive protein (hsCRP), interleukin (IL)-6, and soluble fractions of tumor necrosis factor-α receptors 1 and 2 (sTNFαR1 and sTNFαR2, respectively) were measured. All statistical analyses were stratified by sex.

RESULTS

Subjects with diabetes had a higher aPWV compared with healthy control subjects (men: 6.9 vs. 6.3 m/s, P < 0.001; women: 6.4 vs. 6.0 m/s, P = 0.023). These differences remained significant after adjusting for cardiovascular risk factors. Men with diabetes had higher concentrations of hsCRP (1.2 vs. 0.6 mg/L; P = 0.036), IL-6 (0.6 vs. 0.3 pg/mL; P = 0.002), sTNFαR1 (2,739 vs. 1,410 pg/mL; P < 0.001), and sTNFαR2 (2,774 vs. 2,060 pg/mL; P < 0.001). Women with diabetes only had higher concentrations of IL-6 (0.6 vs. 0.4 pg/mL; P = 0.039). In men with diabetes, aPWV correlated positively with hsCRP (r = 0.389; P = 0.031) and IL-6 (r = 0.447; P = 0.008), whereas in women with diabetes no significant correlation was found. In men, multiple linear regression analysis showed that the following variables were associated independently with aPWV: age, BMI, type 1 diabetes, and low-grade inflammation (R² = 0.543). In women, these variables were age, BMI, mean arterial pressure, and type 1 diabetes (R² = 0.550).

CONCLUSIONS

Arterial stiffness assessed as aPWV is increased in patients with type 1 diabetes without clinical cardiovascular disease, independently of classical cardiovascular risk factors. In men with type 1 diabetes, low-grade inflammation is independently associated with arterial stiffness.It is well established that type 2 diabetes is a risk factor for cardiovascular disease. However, it is less well known that the relative risk of cardiovascular disease in type 1 diabetes can be as much as 10-fold greater than in the healthy population, especially in women (1), being even greater than in type 2 diabetes (2). Consequently, cardiovascular disease is the major cause of mortality in type 1 diabetes (2). Diabetes results in an accelerated arteriosclerotic process, which is not fully explained by classical cardiovascular risk factors. As a result, the pathophysiological mechanisms underlying cardiovascular events in type 1 diabetes are not completely understood.Arterial stiffness is an early sign of arteriosclerosis (3), and its study would be appropriate for investigating the arteriosclerotic mechanisms long before any cardiovascular event occurs. Arterial stiffness predicts cardiovascular events independently of classical cardiovascular risk factors in several populations (see below). Therefore, it can be assumed that it reflects the deleterious effect of all cardiovascular risk factors (known and unknown) on the arterial wall. The gold standard for measuring central arterial stiffness is aortic pulse wave velocity (aPWV), according to a recent consensus (4). aPWV independently predicts cardiovascular events and mortality in the general population, in the elderly, in hypertensive individuals, in subjects with end-stage renal failure, and in subjects with type 2 diabetes (5).Finally, little is known regarding factors involved in the pathophysiology of arterial stiffness in type 1 diabetes. One of these factors could be low-grade inflammation. High-sensitivity C-reactive protein (hsCRP) is the most established downstream marker of low-grade inflammation and has been reported to be a strong predictor of cardiovascular outcomes (6). The primary proinflammatory cytokines interleukin (IL)-6 and tumor necrosis factor (TNF)-α are the main inducers for the hepatic synthesis of hsCRP (7). Although there is less evidence in comparison with hsCRP, both of them also have been associated with the prediction of cardiovascular outcomes (8–10). Low-grade inflammation also has been associated with the development of both micro- and macrovascular complications in type 1 diabetes (11). Indeed, low-grade inflammation impairs endothelial function and has been associated with an increase in aPWV in healthy subjects (12), in hypertensive individuals (13), and in subjects with chronic kidney disease (14) or with metabolic syndrome (15). No such evidence exists in type 1 diabetes; however, recently an activation of the TNFα system has been reported in association with an increase in brachial PP, a subrogate marker of arterial stiffness, in normotensive subjects with type 1 diabetes (16).Our main objective was to evaluate aPWV as a measure of arterial stiffness in a group of subjects with type 1 diabetes without clinical cardiovascular disease and to explore its relationship with biomarkers of low-grade inflammation. Because the role of low-grade inflammation in the atherosclerotic process seems to be different in men and women (17), our study was stratified by sex, and the sample size was calculated taking this stratification into account.     

A human ribonuclease induces apoptosis associated with p21<Superscript>WAF1/CIP1</Superscript>induction and JNK inactivation

Background  

Ribonucleases are promising agents for use in anticancer therapy. Among the different ribonucleases described to be cytotoxic, a paradigmatic example is onconase which manifests cytotoxic and cytostatic effects, presents synergism with several kinds of anticancer drugs and is currently in phase II/III of its clinical trial as an anticancer drug against different types of cancer. The mechanism of cytotoxicity of PE5, a variant of human pancreatic ribonuclease carrying a nuclear localization signal, has been investigated and compared to that of onconase.     

Heterologous prime-boost immunizations with different Salmonella serovars for enhanced antigen-specific CD8 T-cell induction

Pre-existing anti-vector immunity can severely compromise the ability of Salmonella enterica serovar Typhimurium live vaccines to induce protective CD8 T-cell frequencies after type III secretion system-mediated heterologous protein translocation in orally immunized mice. In the present study, we demonstrate that heterologous prime-boost immunizations using attenuated serovar Typhimurium and serovar Dublin strains for foreign antigen delivery can be employed to bypass anti-Salmonella immunity resulting in enhanced antigen-specific CD8 T-cell induction. This desirable effect can be explained by the fact that, in contrast to homologous prime-boost immunizations, vaccination with different Salmonella serovars is characterized by long-lasting colonization of mice by both live carrier vaccines.     

Cell stress induces TDP-43 pathological changes associated with ERK1/2 dysfunction: implications in ALS

TDP-43 has been implicated in the pathogenesis of amyotrophic lateral sclerosis and other neurodegenerative diseases. Here we demonstrate, using neuronal and spinal cord organotypic culture models, that chronic excitotoxicity, oxidative stress, proteasome dysfunction and endoplasmic reticulum stress mechanistically induce mislocalization, phosphorylation and aggregation of TDP-43. This is compatible with a lack of function of this protein in the nucleus, specially in motor neurons. The relationship between cell stress and pathological changes of TDP-43 also includes a dysfunction in the survival pathway mediated by mitogen-activated protein kinase/extracellular signal-regulated kinases (ERK1/2). Thus, under stress conditions, neurons and other spinal cord cells showed cytosolic aggregates containing ERK1/2. Moreover, aggregates of abnormal phosphorylated ERK1/2 were also found in the spinal cord in amyotrophic lateral sclerosis (ALS), specifically in motor neurons with abnormal immunoreactive aggregates of phosphorylated TDP-43. These results demonstrate that cellular stressors are key factors in neurodegeneration associated with TDP-43 and disclose the identity of ERK1/2 as novel players in the pathogenesis of ALS.