Neuron-specific knock-down of SMN1 causes neuron degeneration and death through an apoptotic mechanism.

Spinal muscular atrophy is a devastating disease that is characterized by degeneration and death of a specific subclass of motor neurons in the anterior horn of the spinal cord. Although the gene responsible, survival motor neuron 1 (SMN1), was identified 20 years ago, it has proven difficult to investigate its effects in vivo. Consequently, a number of key questions regarding the molecular and cellular functions of this molecule have remained unanswered. We developed a Caenorhabditis elegans model of smn-1 loss-of-function using a neuron-specific RNA interference strategy to knock-down smn-1 selectively in a subclass of motor neurons. The transgenic animals presented a cell-autonomous, age-dependent degeneration of motor neurons detected as locomotory defects and the disappearance of presynaptic and cytoplasmic fluorescent markers in targeted neurons. This degeneration led to neuronal death as revealed by positive reactivity to genetic and chemical cell-death markers. We show that genes of the classical apoptosis pathway are involved in the smn-1-mediated neuronal death, and that this phenotype can be rescued by the expression of human SMN1, indicating a functional conservation between the two orthologs. Finally, we determined that Plastin3/plst-1 genetically interacts with smn-1 to prevent degeneration, and that treatment with valproic acid is able to rescue the degenerative phenotype. These results provide novel insights into the cellular and molecular mechanisms that lead to the loss of motor neurons when SMN1 function is reduced.

Activating mutations in RRAS underlie a phenotype within the RASopathy spectrum and contribute to leukaemogenesis.

RASopathies, a family of disorders characterized by cardiac defects, defective growth, facial dysmorphism, variable cognitive deficits and predisposition to certain malignancies, are caused by constitutional dysregulation of RAS signalling predominantly through the RAF/MEK/ERK (MAPK) cascade. We report on two germline mutations (p.Gly39dup and p.Val55Met) in RRAS, a gene encoding a small monomeric GTPase controlling cell adhesion, spreading and migration, underlying a rare (2 subjects among 504 individuals analysed) and variable phenotype with features partially overlapping Noonan syndrome, the most common RASopathy. We also identified somatic RRAS mutations (p.Gly39dup and p.Gln87Leu) in 2 of 110 cases of non-syndromic juvenile myelomonocytic leukaemia, a childhood myeloproliferative/myelodysplastic disease caused by upregulated RAS signalling, defining an atypical form of this haematological disorder rapidly progressing to acute myeloid leukaemia. Two of the three identified mutations affected known oncogenic hotspots of RAS genes and conferred variably enhanced RRAS function and stimulus-dependent MAPK activation. Expression of an RRAS mutant homolog in Caenorhabditis elegans enhanced RAS signalling and engendered protruding vulva, a phenotype previously linked to the RASopathy-causing SHOC2(S2G) mutant. Overall, these findings provide evidence of a functional link between RRAS and MAPK signalling and reveal an unpredicted role of enhanced RRAS function in human disease.

LIMITATION OF PRIMERS USED IN PCR FOR THE CHARACTERIZATION OF LEISHMANIA INFANTUM.

Conventional PCR provides Leishmania species characterization with even a small amount of biological material. Species-specific primers have been a widely used alternative; however, nonspecific amplifications are a reality, interfering with PCR efficiency. In endemic areas with multiple etiological agents for leishmaniasis, there is a requirement for higher specificity of primers. This study evaluates 3 pairs of primers described for the identification and characterization of Leishmania infantum. Primers RV1/RV2, LEISH1/LEISH2, and FLC2/RLC2 were used with the DNA of L. infantum, Leishmania amazonensis, and Leishmania braziliensis. An initial temperature curve was performed (52-62 C) to determine the optimal annealing temperature, followed by a dilution curve of Leishmania DNA (500 pg/µl, 50 pg/µl, 5 pg/µl, 500 fg/µl, 50 fg/µl, 5 fg/µl, and 0.5 fg/µl) to be used for analytical sensitivity. RV1/RV2 PCR amplified L. infantum and L. amazonensis at all analyzed temperatures; LEISH1/LEISH2 PCR amplified all 3 species of Leishmania, although at some temperatures L. infantum was specifically amplified, and, finally, FLC2/RLC2 PCR amplified only L. infantum at all temperatures analyzed. In terms of sensitivity, RV1/RV2 PCR detected 1 fg of L. infantum DNA and 100 pg of L. amazonensis DNA; LEISH1/LEISH2 PCR detected 1 fg of L. infantum DNA, 100 fg of L. amazonensis DNA, and 10 fg of L. braziliensis DNA; and FLC2/RLC2 PCR detected 10 fg of L. infantum DNA. Thus, PCR with FLC2/RLC2 primers is best suited for the molecular characterization of L. infantum, especially in areas where there is an incidence of more than 1 Leishmania species, such as South America.

Ozaki Procedure Plus Aortic Annular Enlargement in an Extremely Narrow Aortic Annulus: A Surgical Challenge.

We report a 19-year-old woman with a history of percutaneous aortic valve (AV) balloon valvuloplasty 3 years ago. She was admitted with severe symptomatic AV stenosis with narrow aortic annulus. We decided to perform AV reconstruction with autologous pericardium. However, surgical findings revealed a very narrow AV annulus with a unicuspid valve; therefore, an AV enlargement was added.

[Right intraatrial lipoma. A case report]. / Lipoma intrauricular derecho. Reporte de un caso.

We present the case of a 56-year-old male patient with a history of atrial flutter for six months; he was admitted to the hospital for presenting a mass of 8 cm in maximum diameter in the right atrium, which prolapsed through the tricuspid valve into the right ventricle. Emergency surgery was scheduled, performing exeresis of the tumor and tricuspid annuloplasty. The Pathological anatomy determined that the removed mass corresponded to a cardiac lipoma.

A new surgical technique for left atrial reduction in giant left atrium.

Objective: The study objective was to evaluate the safety and clinical and echocardiographic outcomes of a new surgical technique in adult patients diagnosed with a giant left atrium. Methods: We analyzed a cohort of patients who underwent left atrium reduction surgery between January 2016 and June 2020 performed by a specialized surgical team in 2 national reference centers in Lima, Peru. We assessed the major adverse valvular-related events and the New York Heart Association functional class as primary clinical outcomes. Also, our primary echocardiographic endings were the diameter, area, and volume of the left atrium. We assessed these variables at 3 time periods: baseline (t0), perioperative period (t1), and extended follow-up (t2: 12 ± 3.4 months). We carried out descriptive and bivariate exploratory statistical analysis for dependent measures. Results: We included 17 patients, 70.6% of whom were women. Rheumatic mitral valve disease (76.5%) was the main etiology. We performed 14 (82.4%) mitral valve replacements and 3 repairs. Major adverse valvular-related events occurred in 1 patient (5.9%) (hemorrhagic stroke) at t1. A significant reduction in the size of the left atrium was observed: diameter (77 mm vs 48 mm, P < .001), area (75 cm2 vs 31 cm2, P < .001), and volume (332 cm3 vs 90 cm3, P < .001). Compared with t0 and t1, these echocardiographic findings remained without significant changes during t2. Conclusions: Our surgical left atrium reduction technique was associated with improved clinical functionality and reduced left atrium measures in patients with a giant left atrium undergoing mitral valve surgery.

[Implementing a chronic pain ambulatory care: preliminary results]. / Implementação de atendimento ambulatorial para dor crônica: resultados preliminares.

BACKGROUND AND OBJECTIVES: Pain is one of the most common reason for seeking medical care. This study aimed to analyze patients with chronic pain in Maricá, Rio de Janeiro State, Brazil. METHODS: A transversal retrospective study with 200 patients, who were treated in ambulatory care in a public hospital from June 2014 to December 2015. The variables analyzed were: pain intensity, type of pain, anatomical location, diagnosis and treatment. The data were submitted to statistical analysis, the Fisher's exact test was applied, and the probability p was significant when ≤0.05. RESULTS: We analyzed 200 patients with chronic pain, most of them female (83%). Mean age was 58.6±13.01 years old. The patients were classified in groups by age, six groups with ten years of difference between them. Main age range was the 50-59 years old group, with 49 females (32%) and 5 males (15%). About 65.5% of the total of patients (131) had severe pain (Numeric Rating Sacale was 9.01). Mixed pain was predominant, affecting 108 patients (92 females and 16 males, what represents 55% and 47% of the total of females and males, respectively, that participate in the study). The most prevalent anatomical pain (159 patients, 131 females and 28 males) was in the lower limbs. Lower back pain was present in 113 of the 200 patients (94 females and 19 males). In the 30-39, 50-59, 60-69 years old group, the results for pain locations were significant: p=0.01, p=0.0069, p=0.0003, respectively. CONCLUSION: The prevalence of chronic pain was associated with females in 50-59 years old and severe mixed pain. It was located mainly in lower limbs and lumbar region. The most frequent diagnosis was low back pain followed by fibromyalgia. The patients were informed about their disease and treatment.

Mastoid and opisthion-bimastoid triangles for sex determination in a Brazilian sample / Triangulos mastoideo y opistión-bimastoideo para la determinación del sexo en una muestra brasileña

SUMMARY: Human identification involving mutilation, advanced decomposition, and skeletonized materials is a challenge for professionals in forensic medicine and dentistry. This study was aimed at a morphometric analysis of the mastoid and bimastoid triangles to determine sex in a Brazilian population. The sample included 80 human skulls (34 females and 46 males; age: 18 to 60 years) from individuals with death certificates. Linear measurements (mm) of the mastoid process - right (n=3) and left (n=3) - and the opisthion- bimastoid (n=3) triangles were taken (digital caliper) by two previously trained researchers. Three cranial points - asterion, porion and process - were determined for the right and left mastoid triangle and three others - (1) opisthion and (2) right and (3) left mastoid - for the bimastoid triangle. Heron's formula was used to calculate the area of the triangles assessed. Data were submitted to the Shapiro- Wilk normality test, followed by the unpaired Student's t test or the Mann-Whitney U test, depending on the data distribution verified in the normality test (GraphPad Prism 5.01; p<0.05). The ROC curve was used to measure the accuracy of the variables toward sex determination. The opisthion-bimastoid triangle showed high accuracy and significant differences in all the variables assessed and thus was considered a potential element for sex determination in the Brazilian population assessed.

RESUMEN: La identificación humana que comprende la mutilación, descomposición avanzada y materiales esqueletizados, es un desafío para los profesionales de la medicina forense y de la odontología. Este estudio tuvo como objetivo un análisis morfométrico de los triángulos mastoideo y bimastoideo para determinar el sexo en una población brasileña. La muestra incluyó 80 cráneos humanos (34 mujeres y 46 hombres; edad: 18 a 60 años) de sujetos con certificado de defunción. Se tomaron las medidas lineales (mm) del proceso mastoides - derecha (n = 3) e izquierda (n = 3) - y los triángulos opistion-bimastoide (n = 3) (caliper digital) por dos investigadores previamente entrenados. Se determinaron tres puntos craneales - asterion, porion y proceso mastoides - para el triángulo mastoideo derecho e izquierdo y otros tres - (1) opistion y (2) proceso mastoides derecho e izquierdo - para el triángulo bimastoideo. Se utilizó la fórmula de Heron para calcular el área de los triángulos evaluados. Los datos se analizaron con la prueba de normalidad de Shapiro-Wilk, seguida de la prueba t de Student para datos no apareados o la prueba U de Mann-Whitney, según la distribución de datos verificada en la prueba de normalidad (GraphPad Prism 5.01; p <0.05). La curva ROC se utilizó para medir la precisión de las variables hacia la determinación del sexo. El triángulo opistion-bimastoide mostró una alta precisión y diferencias significativas en todas las variables evaluadas, por lo que se consideró un elemento probable para la determinación del sexo en la población brasileña evaluada.

Adherence to heroin dependence therapies and human immunodeficiency virus/acquired immunodeficiency syndrome infection rates among drug abusers.

Adherence is a primary determinant of treatment effectiveness; thus, poor adherence attenuates optimum clinical benefit. A bibliographic review was conducted to evaluate the impact of adherence to heroin dependence treatment on human immunodeficiency virus (HIV) transmission and to identify interventions proven to be effective in improving adherence. The best adherence rates were achieved by methadone and diacetylmorphine, both of which are comparable in promoting significant reduction in heroin use. Methadone adjusted-dose studies with daily doses ranging from 100 to 200 mg and multiple support interventions achieved the highest adherence rates. Studies of methadone maintenance that examined changes in HIV prevalence of infection have found that higher treatment adherence is correlated with a reduction in HIV transmission. These data suggest that patients who adhere continuously to methadone treatment are less likely to continue injecting illicit drugs and sharing contaminated injection equipment than are those who interrupt treatment, thus preventing the spread of HIV via drug injection.

HPLC determination of amoxicillin comparative bioavailability in healthy volunteers after a single dose administration.

PURPOSE: An accurate, precise and sensitive HPLC assay was developed for the determination of amoxicillin in human plasma samples, to compare the bioavailability of two amoxicillin capsule (500mg) formulations (Amoxicilina from Brazil, as a test formulation and Amoxil from SmithKline Beecham Laboratories Ltda., Brazil, as a reference formulations) in 24 volunteers of both sexes. METHODS: Amoxicillin concentrations were analyzed by combined reversed phase liquid chromatography and UV detection (lambda=229 nm). Amoxicillin and cefadroxil (internal standard) were extracted from the plasma by addition of cold methanol. The separation was achieved using the Lichrosorb 10 microm, C18 reversed phase column at room temperature. The mobile phase consisted of a 95% phosphate buffer (0.01 mol/L), pH=4.8 and 5% acetonitrile mixture. The study was conducted using an open randomized 2-period crossover balanced design with a 1-week washout period between the doses. Plasma samples were obtained over an 8-hour period. The bioequivalence between the two formulations was assessed by calculating individual peak plasma concentrations (C(max) ) and area under the curve (AUC(0-8h) ) ratios (test/reference). The statistical interval proposed was 80-125%, as established by the US Food and drug administration Agency. RESULTS: The internal standard and amoxicillin eluted about 4.2 and 5.2 min, respectively at a flow rate of 1.3ml/min. The mean absolute recovery of AMO in plasma was 90.0% at 3 microg/ml, 98.6% at 25 microg/ml and 95.3 at 50 microg/ml. The assay showed excellent relationships between peak height ratios and plasma concentrations (r(2)>or= 0.999). The limit of quantification was 1g/ml, based on 200l of plasma. The geometric mean of Amoxicilina/Amoxil 500 mg capsules individual percentage ratio was 101.4% for AUC(0-8h), and 99.9% for C(max). The 90% confidence intervals were 98.3-104.4% and 95.7-103.9%, respectively. CONCLUSION: This simple, rapid and selective method is suitable for pharmacokinetic, bioavailability and bioequivalence studies. Since the 90% CI for both C(max )and AUC(0-8h) lies within the 80-125% interval proposed by the Food and Drug Administration, it was concluded that Amoxicilina 500 mg capsules was bioequivalent to Amoxil capsules 500 mg, in terms of both the rate and extent of absorption.

Atenolol quantification in human plasma by high-performance liquid chromatography: application to bioequivalence study.

An accurate, precise, and sensitive high-performance liquid chromatography (HPLC) assay was developed for the determination of atenolol in human plasma samples to compare the bioavailability of 2 atenolol tablet (50 mg) formulations in 24 volunteers of both sexes. The study had an open, randomized, 2-period crossover design with a 1-week washout period. Plasma samples were obtained over a 24-hour interval. Atenolol concentrations were analyzed by combined reversed phase liquid chromatography and fluorescence detection (lambda(EX) = 258 nm, lambda(EM) = 300 nm). From the atenolol plasma concentration versus time curves, the following pharmacokinetic parameters were obtained: AUC(0-24h), AUC(0- infinity ), and C(max). The geometric mean of test/reference 50-mg tablets individual percent ratio was 102.2% for AUC(0-24h), and 101.6% for C(max). The 90% confidence intervals (CI) were 100.2% to 105.4% and 100.9% to 103.5%, respectively. Since the 90% CI for both C(max) and AUC(0-24h) were within the 80% to 125% interval proposed by the Food and Drug Administration, it was concluded that atenolol (50-mg tablets) test formulation was bioequivalent to the reference formulation, with regard to both the rate and extent of absorption.

Implementing a chronic pain ambulatory care: preliminary results / Implementação de atendimento ambulatorial para dor crônica: resultados preliminares

Abstract Background and objectives: Pain is one of the most common reason for seeking medical care. This study aimed to analyze patients with chronic pain in Maricá, Rio de Janeiro State, Brazil. Methods: A transversal retrospective study with 200 patients, who were treated in ambulatory care in a public hospital from June 2014 to December 2015. The variables considered were: pain intensity, type of pain, anatomical location, diagnosis and treatment. The data were statistically analyzed, the Fisher's exact test was applied, and the probability p was significant when ≤0.05. Results: We analyzed 200 patients with chronic pain, most of them female (83%). Mean age was 58.6 ± 13.01 years old. The patients were classified in groups by age, six groups with ten years of difference between them. Main age range was the 50-59 years old group, with 49 females (32%) and 5 males (15%). About 65.5% of the total of patients (131) had severe pain (Numeric Rating Sacale was 9.01). Mixed pain was predominant, affecting 108 patients (92 females and 16 males, what represents 55% and 47% of the total of females and males, respectively, that participate in the study). The most prevalent anatomical pain (159 patients, 131 females and 28 males) was in the lower limbs. Lower back pain was present in 113 of the 200 patients (94 females and 19 males). In the 30-39, 50-59, 60-69 years old group, the results for pain locations were significant: p = 0.01, p = 0.0069, p = 0.0003, respectively. Conclusion: The prevalence of chronic pain was associated with females in 50-59 years old and severe mixed pain. Pain was located mainly in lower limbs and lumbar region. The most frequent diagnosis was low back pain followed by fibromyalgia. The patients were informed about their disease and treatment.

Resumo Justificativa e objetivos: A dor é um dos motivos mais comuns para procurar atendimento médico. Este estudo teve como objetivo analisar pacientes com dor crônica atendidos em Maricá, no Estado do Rio de Janeiro, Brasil. Métodos: Estudo transversal retrospectivo com 200 pacientes, atendidos em ambulatório de um hospital público no período de junho de 2014 a dezembro de 2015. As variáveis analisadas foram: intensidade da dor, tipo de dor, localização anatômica, diagnóstico e tratamento. Os dados foram submetidos à análise estatística, aplicou-se o teste exato de Fisher, e o valor p foi significativo quando ≤ 0,05. Resultados: Analisamos 200 pacientes com dor crônica, sendo a maioria mulheres (83%). A média de idade foi de 58,6 ± 13,01 anos. Os pacientes foram classificados em seis grupos de acordo com a faixa etária, com dez anos de diferença entre eles. O grupo principal foi entre 50-59 anos, com 49 mulheres (32%) e cinco homens (15%). Dos pacientes, 65,5% apresentaram dor intensa (Escala Visual Numérica 9,01). A dor mista foi prevalente, afetou 108 pacientes (92 mulheres e 16 homens, o que representa 55% e 47% do total de mulheres e homens, respectivamente). A dor anatômica mais prevalente (159 pacientes, 131 mulheres e 28 homens) foi nos membros inferiores. A dor na parte inferior das costas estava presente em 113 das 200 pessoas analisadas (94% mulheres e 19% homens). Nos grupos entre 30-39, 50-59 e 60-69 anos, os resultados para a localização da dor foram significativos: p = 0,01, p = 0,0069, p = 0,0003, respectivamente. Conclusão: A prevalência de dor crônica foi associada ao sexo feminino na faixa de 50-59 anos e à dor mista intensa. A dor foi localizada principalmente nos membros inferiores e na região lombar. O diagnóstico mais frequente foi de lombalgia seguida de fibromialgia. Os pacientes foram informados sobre suas doenças e tratamento.

Performance of pilot-scale constructed wetlands for secondary treatment of chromium-bearing tannery wastewaters.

Tannery operations consist of converting raw animal skins into leather through a series of complex water- and chemically-intensive batch processes. Even when conventional primary treatment is supplemented with chemicals, the wastewater requires some form of biological treatment to enable the safe disposal to the natural environment. Thus, there is a need for the adoption of low cost, reliable, and easy-to-operate alternative secondary treatment processes. This paper reports the findings of two pilot-scale wetlands for the secondary treatment of primary effluents from a full tannery operation in terms of resilience (i.e., ability to produce consistent effluent quality in spite of variable influent loads) and reliability (i.e., ability to cope with sporadic shock loads) when treating this hazardous effluent. Areal mass removal rates of 77.1 g COD/m2/d, 11 g TSS/m2/d, and 53 mg Cr/m2/d were achieved with a simple gravity-flow horizontal subsurface flow unit operating at hydraulic loading rates of as much as 10 cm/d. Based on the findings, a full-scale wetland was sized to treat all the effluent from the tannery requiring 68% more land than would have been assumed based on literature values. Constructed wetlands can offer treatment plant resilience for minimum operational input and reliable effluent quality when biologically treating primary effluents from tannery operations.