Localization of Secondary Metabolites in Marine Invertebrates: Contribution of MALDI MSI for the Study of Saponins in Cuvierian Tubules of H. forskali

BACKGROUND: Several species of sea cucumbers of the family Holothuriidae possess a particular mechanical defense system called the Cuvierian tubules (Ct). It is also a chemical defense system as triterpene glycosides (saponins) appear to be particularly concentrated in Ct. In the present study, the precise localization of saponins in the Ct of Holothuria forskali is investigated. Classical histochemical labeling using lectin was firstly performed but did not generate any conclusive results. Thus, MALDI mass spectrometry Imaging (MALDI-MSI) was directly applied and completed by statistical multivariate tests. A comparison between the tubules of relaxed and stressed animals was realized. RESULTS: These analyses allowed the detection of three groups of ions, corresponding to the isomeric saponins of the tubules. Saponins detected at m/z 1287 and 1303 were the most abundant and were apparently localized in the connective tissue of the tubules of both relaxed and stressed individuals. Saponins at m/z 1125 and 1141 were detected in lower amount and were present in tissues of relaxed animals. Finally, saponin ions at 1433, 1449, 1463 and 1479 were observed in some Ct of stressed holothuroids in the outer part of the connective tissue. The saponin group m/z 14xx seems therefore to be stress-specific and could originate from modifications of the saponins with m/z of 11xx. CONCLUSIONS: All the results taken together indicate a complex chemical defense mechanism with, for a single organ, different sets of saponins originating from different cell populations and presenting different responses to stress. The present study also reflects that MALDI-MSI is a valuable tool for chemical ecology studies in which specific chemical signalling molecules like allelochemicals or pheromones have to be tracked. This report represents one of the very first studies using these tools to provide a functional and ecological understanding of the role of natural products from marine invertebrates

Transcriptional Responses of Cultured Rat Sympathetic Neurons during BMP-7-Induced Dendritic Growth

Dendrites are the primary site of synapse formation in the vertebrate nervous system; however, relatively little is known about the molecular mechanisms that regulate the initial formation of primary dendrites. Embryonic rat sympathetic neurons cultured under defined conditions extend a single functional axon, but fail to form dendrites. Addition of bone morphogenetic proteins (BMPs) triggers these neurons to extend multiple dendrites without altering axonal growth or cell survival. We used this culture system to examine differential gene expression patterns in naïve vs. BMP-treated sympathetic neurons in order to identify candidate genes involved in regulation of primary dendritogenesis.To determine the critical transcriptional window during BMP-induced dendritic growth, morphometric analysis of microtubule-associated protein (MAP-2)-immunopositive processes was used to quantify dendritic growth in cultures exposed to the transcription inhibitor actinomycin-D added at varying times after addition of BMP-7. BMP-7-induced dendritic growth was blocked when transcription was inhibited within the first 24 hr after adding exogenous BMP-7. Thus, total RNA was isolated from sympathetic neurons exposed to three different experimental conditions: (1) no BMP-7 treatment; (2) treatment with BMP-7 for 6 hr; and (3) treatment with BMP-7 for 24 hr. Affymetrix oligonucleotide microarrays were used to identify differential gene expression under these three culture conditions. BMP-7 significantly regulated 56 unique genes at 6 hr and 185 unique genes at 24 hr. Bioinformatic analyses implicate both established and novel genes and signaling pathways in primary dendritogenesis.This study provides a unique dataset that will be useful in generating testable hypotheses regarding transcriptional control of the initial stages of dendritic growth. Since BMPs selectively promote dendritic growth in central neurons as well, these findings may be generally applicable to dendritic growth in other neuronal cell types

Effect of SGLT2 inhibitors on stroke and atrial fibrillation in diabetic kidney disease: Results from the CREDENCE trial and meta-analysis

BACKGROUND AND PURPOSE: Chronic kidney disease with reduced estimated glomerular filtration rate or elevated albuminuria increases risk for ischemic and hemorrhagic stroke. This study assessed the effects of sodium glucose cotransporter 2 inhibitors (SGLT2i) on stroke and atrial fibrillation/flutter (AF/AFL) from CREDENCE (Canagliflozin and Renal Events in Diabetes With Established Nephropathy Clinical Evaluation) and a meta-Analysis of large cardiovascular outcome trials (CVOTs) of SGLT2i in type 2 diabetes mellitus. METHODS: CREDENCE randomized 4401 participants with type 2 diabetes mellitus and chronic kidney disease to canagliflozin or placebo. Post hoc, we estimated effects on fatal or nonfatal stroke, stroke subtypes, and intermediate markers of stroke risk including AF/AFL. Stroke and AF/AFL data from 3 other completed large CVOTs and CREDENCE were pooled using random-effects meta-Analysis. RESULTS: In CREDENCE, 142 participants experienced a stroke during follow-up (10.9/1000 patient-years with canagliflozin, 14.2/1000 patient-years with placebo; hazard ratio [HR], 0.77 [95% CI, 0.55-1.08]). Effects by stroke subtypes were: ischemic (HR, 0.88 [95% CI, 0.61-1.28]; n=111), hemorrhagic (HR, 0.50 [95% CI, 0.19-1.32]; n=18), and undetermined (HR, 0.54 [95% CI, 0.20-1.46]; n=17). There was no clear effect on AF/AFL (HR, 0.76 [95% CI, 0.53-1.10]; n=115). The overall effects in the 4 CVOTs combined were: Total stroke (HRpooled, 0.96 [95% CI, 0.82-1.12]), ischemic stroke (HRpooled, 1.01 [95% CI, 0.89-1.14]), hemorrhagic stroke (HRpooled, 0.50 [95% CI, 0.30-0.83]), undetermined stroke (HRpooled, 0.86 [95% CI, 0.49-1.51]), and AF/AFL (HRpooled, 0.81 [95% CI, 0.71-0.93]). There was evidence that SGLT2i effects on total stroke varied by baseline estimated glomerular filtration rate (P=0.01), with protection in the lowest estimated glomerular filtration rate (45 mL/min/1.73 m2]) subgroup (HRpooled, 0.50 [95% CI, 0.31-0.79]). CONCLUSIONS: Although we found no clear effect of SGLT2i on total stroke in CREDENCE or across trials combined, there was some evidence of benefit in preventing hemorrhagic stroke and AF/AFL, as well as total stroke for those with lowest estimated glomerular filtration rate. Future research should focus on confirming these data and exploring potential mechanisms

Canagliflozin and Renal Outcomes in Type 2 Diabetes and Nephropathy

BACKGROUND Type 2 diabetes mellitus is the leading cause of kidney failure worldwide, but few effective long-term treatments are available. In cardiovascular trials of inhibitors of sodium–glucose cotransporter 2 (SGLT2), exploratory results have suggested that such drugs may improve renal outcomes in patients with type 2 diabetes. METHODS In this double-blind, randomized trial, we assigned patients with type 2 diabetes and albuminuric chronic kidney disease to receive canagliflozin, an oral SGLT2 inhibitor, at a dose of 100 mg daily or placebo. All the patients had an estimated glomerular filtration rate (GFR) of 30 to 300 to 5000) and were treated with renin–angiotensin system blockade. The primary outcome was a composite of end-stage kidney disease (dialysis, transplantation, or a sustained estimated GFR of <15 ml per minute per 1.73 m 2), a doubling of the serum creatinine level, or death from renal or cardiovascular causes. Prespecified secondary outcomes were tested hierarchically. RESULTS The trial was stopped early after a planned interim analysis on the recommendation of the data and safety monitoring committee. At that time, 4401 patients had undergone randomization, with a median follow-up of 2.62 years. The relative risk of the primary outcome was 30% lower in the canagliflozin group than in the placebo group, with event rates of 43.2 and 61.2 per 1000 patient-years, respectively (hazard ratio, 0.70; 95% confidence interval [CI], 0.59 to 0.82; P=0.00001). The relative risk of the renal-specific composite of end-stage kidney disease, a doubling of the creatinine level, or death from renal causes was lower by 34% (hazard ratio, 0.66; 95% CI, 0.53 to 0.81; P<0.001), and the relative risk of end-stage kidney disease was lower by 32% (hazard ratio, 0.68; 95% CI, 0.54 to 0.86; P=0.002). The canagliflozin group also had a lower risk of cardiovascular death, myocardial infarction, or stroke (hazard ratio, 0.80; 95% CI, 0.67 to 0.95; P=0.01) and hospitalization for heart failure (hazard ratio, 0.61; 95% CI, 0.47 to 0.80; P<0.001). There were no significant differences in rates of amputation or fracture. CONCLUSIONS In patients with type 2 diabetes and kidney disease, the risk of kidney failure and cardiovascular events was lower in the canagliflozin group than in the placebo group at a median follow-up of 2.62 years

Canagliflozin and renal outcomes in type 2 diabetes and nephropathy

BACKGROUND Type 2 diabetes mellitus is the leading cause of kidney failure worldwide, but few effective long-term treatments are available. In cardiovascular trials of inhibitors of sodium–glucose cotransporter 2 (SGLT2), exploratory results have suggested that such drugs may improve renal outcomes in patients with type 2 diabetes. METHODS In this double-blind, randomized trial, we assigned patients with type 2 diabetes and albuminuric chronic kidney disease to receive canagliflozin, an oral SGLT2 inhibitor, at a dose of 100 mg daily or placebo. All the patients had an estimated glomerular filtration rate (GFR) of 30 to &lt;90 ml per minute per 1.73 m2 of body-surface area and albuminuria (ratio of albumin [mg] to creatinine [g], &gt;300 to 5000) and were treated with renin–angiotensin system blockade. The primary outcome was a composite of end-stage kidney disease (dialysis, transplantation, or a sustained estimated GFR of &lt;15 ml per minute per 1.73 m2), a doubling of the serum creatinine level, or death from renal or cardiovascular causes. Prespecified secondary outcomes were tested hierarchically. RESULTS The trial was stopped early after a planned interim analysis on the recommendation of the data and safety monitoring committee. At that time, 4401 patients had undergone randomization, with a median follow-up of 2.62 years. The relative risk of the primary outcome was 30% lower in the canagliflozin group than in the placebo group, with event rates of 43.2 and 61.2 per 1000 patient-years, respectively (hazard ratio, 0.70; 95% confidence interval [CI], 0.59 to 0.82; P=0.00001). The relative risk of the renal-specific composite of end-stage kidney disease, a doubling of the creatinine level, or death from renal causes was lower by 34% (hazard ratio, 0.66; 95% CI, 0.53 to 0.81; P&lt;0.001), and the relative risk of end-stage kidney disease was lower by 32% (hazard ratio, 0.68; 95% CI, 0.54 to 0.86; P=0.002). The canagliflozin group also had a lower risk of cardiovascular death, myocardial infarction, or stroke (hazard ratio, 0.80; 95% CI, 0.67 to 0.95; P=0.01) and hospitalization for heart failure (hazard ratio, 0.61; 95% CI, 0.47 to 0.80; P&lt;0.001). There were no significant differences in rates of amputation or fracture. CONCLUSIONS In patients with type 2 diabetes and kidney disease, the risk of kidney failure and cardiovascular events was lower in the canagliflozin group than in the placebo group at a median follow-up of 2.62 years

Aspectos fisiopatológicos del envejecimiento humano y caídas en adultos mayores

Made available in DSpace on 2015-06-22T12:26:40Z (GMT). No. of bitstreams: 2 license.txt: 1914 bytes, checksum: 7d48279ffeed55da8dfe2f8e81f3b81f (MD5) sandra_silvaetal_IOC_2014.pdf: 188647 bytes, checksum: d1b966ca79e842af51790c31fe48957c (MD5) Previous issue date: 2014Universidade do Estado do Rio de Janeiro (UERJ). Faculdade de Ciências Médicas. Departamento de Patologia e Laboratórios. Rio de Janeiro, RJ, Brasil.Universidade do Estado do Rio de Janeiro (UERJ). Faculdade de Ciências Médicas. Departamento de Patologia e Laboratórios. Rio de Janeiro, RJ, Brasil.Universidade do Estado do Rio de Janeiro (UERJ). Faculdade de Ciências Médicas. Departamento de Patologia e Laboratórios. Rio de Janeiro, RJ, Brasil.As quedas são um problema de saúde pública entre os idosos, em vista da mortalidade, morbidade e dos custos social e econômico. O tema é muito valorizado pela gerontologia e uma fonte de preocupação para os pesquisadores dessa área, principalmente quando pessoas denominam esse evento como sendo normal e próprio do processo de envelhecimento. Queda pode ser definida como um evento não intencional que tem como resultado a mudança de posição do indivíduo para um nível mais baixo, em relação a sua posição inicial. Pessoas de todas as idades apresentam risco de sofrer queda. Porém, para os idosos, elas possuem um significado muito relevante, pois podem levá-lo à incapacidade, injúria e morte. Seu custo social é imenso e torna-se maior quando o idoso tem diminuição da autonomia e da independência ou passa a necessitar de institucionalização. Sabe-se que é elevado o número de idosos que caem e que mudam radicalmente sua vida cotidiana, tanto pela queda em si, como pelo temor de uma nova ocorrência, restringindo suas atividades, aumentando o isolamento social e o declínio na saúde, repercutindo no risco de serem institucionalizados. Dentre os fatores de risco, aqueles de maior ocorrência são a deterioração da visão, uso simultâneo de medicamentos (especialmente diuréticos e psicoativos) e flexibilidade reduzida (quadril e tornozelos), fatores estes que deveriam ser considerados em programas para prevenção de quedas em idosos. Além de prejuízo físico e psicológico, esses acidentes geram um aumento dos custos com cuidados de saúde, expressos pela utilização de serviços especializados e aumento de hospitalizações. Assim, são numerosos os estudos que analisam as quedas em idosos, tanto no que diz respeito à epidemiologia, etiologia e fatores de risco associados, como em relação às consequências desses eventos. O presente estudo teve como objetivo fazer uma revisão sobre o histórico de quedas em idosos e sugerir de que forma a fisioterapia interferirá para minimizar a ocorrência das mesmas.The natural aging process involves several biological changes that are inherent to organisms and occur on a gradual basis, due to evolutionary needs. This study focuses on the way age-related anatomical and physiological alterations are closely related to the risk of falls in the elderly. Initially, we show how these alterations, which start at the beginning of adult life, shall only acquire, due to the redundant composition of organic systems, relevance and functional significance when the decline either reaches a considerable extension or is associated with the emergence of pathologies. The speed of such a decline depends on several factors, both genetic and epigenetic ones, which shall determine the organism’s response to stimuli. Our goal was to analyze the organic systems closely associated with the risk of falls in physiological aging, such as visual and vestibular system, central nervous system (CNS), cardiovascular system and musculoskeletal and bone systems. Thus, we initially addressed the factors responsible for a reduction in the ability to keep stability and posture, the ability to transpose obstacles, visual acuity, and vestibular function. In particular, we highlighted the alterations of the musculoskeletal and bone systems, which are responsible for locomotion and flexibility. Age-related atrophy and muscular weakness may lead to sarcopenia, a syndrome with serious consequences for the elderly, which results in a significant number of falls and fractures. We also discuss how both structural and functional physiological modifications of the cardiovascular system during aging act as adaptive mechanisms for the compensation of further overload. Some extrinsic factors, such as the use of diuretic and antihypertensive drugs, which are often administrated to elderly affected by cardiovascular diseases, also influence the compromised postural stability, thus contributing to the high prevalence of falls in this population, possibly with serious consequences, including death. Finally, we analyze the neurological alterations observed during life emphasizing aspects related to the system which maintains human balance. In association with motor disturbances, such as in strength and balance, these changes use to lead to disabilities and offer a higher risk of morbidity and mortality among elderly, particularly when those result from falls.El proceso natural del envejecimiento implica numerosas transformaciones biológicas inherentes a los organismos, que ocurren de manera gradual e impulsada por necesidades evolutivas. Este artículo aborda cómo las alteraciones anatómicas y fisiológicas propias del envejecimiento están estrechamente relacionadas con el riesgo de caídas en adultos mayores. Comenzamos mostrando cómo esas modificaciones, que se dan en el inicio de la vida adulta, debido a la composición redundante de los sistemas orgánicos, sólo se vuelven importantes y funcionalmente significativas cuando el declive alcanza una extensión considerable o se asocia a la aparición de patologías. La velocidad de este declive depende de varios factores, genéticos y epigenéticos, que determinarán la respuesta del organismo a los estímulos. Intentamos analizar los sistemas orgánicos más concernidos con el riesgo de caídas por el envejecimiento fisiológico, tales como: sistema visual y vestibular; sistema nervioso central (SNC) y cardiovascular; sistema músculo esquelético y óseo. De esta forma, tratamos primeramente los factores responsables por la disminución de la capacidad para mantener la estabilidad y la postura, la transposición de obstáculos, la acuidad visual y la función vestibular. Se hizó énfasis en las alteraciones de los sistemas músculo esqueléticos y óseo, responsables por la locomoción y flexibilidad del cuerpo. La atrofia y la debilidad muscular propias de la edad pueden llevar a la sarcopenia, un síndrome con graves consecuencias para los adultos mayores y responsable por un número substancial de caídas y fracturas. También discutimos cómo las modificaciones fisiológicas estructurales y funcionales del sistema cardiovascular, que se dan en el envejecimiento, actúan como mecanismos adaptativos compensatorios en las situaciones de sobrecarga. Factores extrínsecos, como el uso de medicamentos diuréticos y antihipertensivos administrados con frecuencia en adultos mayores con enfermedades cardiovasculares, también influyen en el déficit de estabilidad postural, contribuyendo a la alta prevalencia de caídas en esta población, pudiendo causar serias consecuencias, incluso la muerte. Finalmente, analizamos las alteraciones neurológicas observadas a lo largo de la vida, enfatizando en los aspectos que se refieren al sistema que mantiene el equilibrio humano. Al asociarlas a transtornos motores, como fuerza y equilibrio, tales alteraciones muchas veces son incapacitantes y presentan un mayor riesgo de morbimortalidad en la población adulta mayor, sobre todo si fueron resultantes de caídas

A frequência do HPV na mucosa oral normal de indivíduos sadios por meio da PCR The frequency of human papillomavirus findings in normal oral mucosa of healthy people by PCR

Os papilomavírus humanos (HPV) são DNA vírus pertencentes à família papilomaviridae com grupos de baixo e alto risco que infectam a pele e a mucosa podendo induzir a formação de tumores epiteliais benignos e malignos. Na mucosa oral, estes vírus têm sido associados a papilomas orais, hiperplasias epiteliais focais, leucoplasias e neoplasias orais. OBJETIVO: Estudar a frequência do HPV em mucosa oral de indivíduos normais. MATERIAL E MÉTODO: Trabalho prospectivo em coorte transversal. Participaram desse estudo 100 indivíduos voluntários, faixa etária de 20 a 31 anos, estudantes universitários, sem história, queixas ou lesões visíveis ao exame físico de cavidade oral e orofaringe. Foram submetidos a questionário com perguntas referentes à epidemiologia da infecção pelo HPV. Foi colhido material de mucosa oral por raspado com escova e analisado pelo PCR. RESULTADOS: Os resultados mostraram ausência de HPV em todas as amostras. CONCLUSÃO: Parece ter havido participação do alto nível socioeconômico com alimentação rica em carotenoides e vitamina C, baixo consumo tabágico e etílico e comportamento heterossexual predominantemente monogâmico com uso regular de preservativos.<br>The human papillomavirus (HPV) is a DNA virus, which belongs to papillomaviridae family, being of low and high risk, which infect the skin and mucous membranes and can induce benign and malign tumor formation. In the oral mucosa they have been associated with oral papilloma, focal epithelial hyperplasia, leucoplakia and oral neoplasia. AIM: to study the frequency of HPV finding in oral mucosa of normal people. MATERIALS AND METHODS: Prospective study, cross-sectional cohort. One hundred volunteers, young adults, healthy, aged between 20 and 31 years, university students with no history, no complains, without oral or oropharyngeal lesions. They were submitted to a questionnaire with questions regarding HPV infection epidemiology. The samples were harvested by brushing and analyzed by PCR. RESULTS: The results were negative for HPV in all samples. CONCLUSION: It seems we had high social and economical class individuals, with nutrition rich in carotenoyds and vitamin C, low smoking and alcohol consumption and heterosexual habits with predominant monogamy and regular use of condoms