Overexpression SPRY2 Suppresses Esophageal Squamous Cell Carcinoma Progression Via Inactivation of ERK/AKT Signaling / La Sobreexpresión SPRY2 Suprime la Evolución del Carcinoma de Células Escamosas de Esófago Mediante la Inactivación de la Señalización ERK/AKT

SUMMARY: Esophageal cancer is one of the most aggressive gastrointestinal cancers. Invasion and metastasis are the main causes of poor prognosis of esophageal cancer. SPRY2 has been reported to exert promoting effects in human cancers, which controls signal pathways including PI3K/AKT and MAPKs. However, the expression of SPRY2 in esophageal squamous cell carcinoma (ESCC) and its underlying mechanism remain unclear. In the present study, we aimed to investigate the detailed role of SPRY2 in the regulation of cell proliferation, invasion and ERK/AKT signaling pathway in ESCC. It was identified that the expression level of SPRY2 in ESCC was remarkably decreased compared with normal tissues, and it was related to clinicopathologic features and prognosis ESCC patients. The upregulation of SPRY2 expression notably inhibited the proliferation, migration and invasion of Eca-109 cells. In addition, the activity of ERK /AKT signaling was also suppressed by the SPRY2 upregulation in Eca-109 cells. Our study suggests that overexpression of SPRY2 suppress cancer cell proliferation and invasion of by through suppression of the ERK/AKT signaling pathways in ESCC. Therefore, SPRY2 may be a promising prognostic marker and therapeutic target for ESCC.

El cáncer de esófago es uno de los cánceres gastrointestinales más agresivos. La invasión y la metástasis son las principales causas de mal pronóstico del cáncer de esófago. Se ha informado que SPRY2 ejerce efectos promotores en los cánceres humanos, que controla las vías de señales, incluidas PI3K/AKT y MAPK. Sin embargo, la expresión de SPRY2 en el carcinoma de células escamosas de esófago (ESCC) y su mecanismo subyacente aún no están claros. En el presente estudio, nuestro objetivo fue investigar el papel detallado de SPRY2 en la regulación de la proliferación celular, la invasión y la vía de señalización ERK/AKT en ESCC. Se identificó que el nivel de expresión de SPRY2 en ESCC estaba notablemente disminuido en comparación con los tejidos normales, y estaba relacionado con las características clínico-patológicas y el pronóstico de los pacientes con ESCC. La regulación positiva de la expresión de SPRY2 inhibió notablemente la proliferación, migración e invasión de células Eca-109. Además, la actividad de la señalización de ERK/AKT también fue suprimida por la regulación positiva de SPRY2 en las células Eca-109. Nuestro estudio sugiere que la sobreexpresión de SPRY2 suprime la proliferación y la invasión de células cancerosas mediante la supresión de las vías de señalización ERK/AKT en ESCC. Por lo tanto, SPRY2 puede ser un marcador de pronóstico prometedor y un objetivo terapéutico para la ESCC.

Narcolepsy: the impact of aging, hypocretin deficiency, and years of formal education in olfactory function and abdominal obesity / Narcolepsia: o impacto da idade, deficiência de hipocretina e anos de estudo na função olfatória e na obesidade central

ABSTRACT Background: Narcolepsy is a disease resulting from the loss of hypocretin-producing cells or other dysfunctions of the hypocretinergic system. In addition to sleep disorders, affected patients may experience increased weight gain, olfactory changes, and poorer quality of life. Methods: This study aimed to investigate the relationship between narcolepsy and weight gain, years of study, sleep parameters, and olfactory dysfunction in patients with narcolepsy type 1 and narcolepsy type 2. Anthropometric, olfactory, socioeducational, and excessive daytime sleepiness evaluations were performed in 77 patients. Results: Greater weight gain and abdominal obesity were observed in patients with type 1 narcolepsy. Patients with higher education level had lower scores of daytime sleepiness, higher scores on the olfactory function test, and lower rates of abdominal obesity. Discussion: Patients with narcolepsy type 1 showed an increased body weight and abdominal obesity when compared to narcolepsy type 2. The patients with a higher schooling level showed a reduction of the daytime sleepiness scores, lower rates of abdominal obesity, and better scores on the olfactory function test. Conclusion: Among all the patients with narcolepsy, the data indicated that aging and hypocretin deficiency are associated with abdominal obesity, while years of study is the variable that mostly influences olfaction function.

RESUMO Antecedentes: A narcolepsia é resultante da perda de células produtoras de hipocretina ou da disfunção do sistema hipocretinérgico. Além dos distúrbios do sono característicos da doença, os pacientes afetados podem apresentar também aumento de peso, alterações olfatórias e pior qualidade de vida. Métodos: O objetivo do estudo é investigar a relação entre a narcolepsia e o ganho de peso, anos de estudo, parâmetros do sono e a disfunção olfatória em pacientes com narcolepsia tipo 1 e narcolepsia tipo 2. Foram realizadas avaliações antropométricas, do olfato, sociais, educacionais e da sonolência excessiva diurna nos 77 indivíduos participantes da pesquisa. Resultados: Foram observados, nos pacientes com narcolepsia tipo 1, maior ganho de peso e maior frequência de obesidade central. Pacientes com ensino superior apresentaram escores mais baixos de sonolência excessiva diurna, escores mais altos no teste de função olfatória e menores taxas de obesidade central. Discussão: Pacientes com narcolepsia tipo 1 apresentaram maior ganho de peso e obesidade central quando comparados aos com narcolepsia tipo 2. Os pacientes com maior escolaridade apresentaram menores escores de sonolência diurna, de obesidade central e melhores escores no teste da função olfatória. Conclusão: Nos indivíduos com narcolepsia tipo 1 e tipo 2, os dados indicaram que o envelhecimento e a deficiência de hipocretina estão associados à obesidade central, enquanto anos de estudo é a variável que mais influencia na função olfatória.

Apoplexy in sporadic pituitary adenomas: a single referral center experience and AIP mutation analysis

ABSTRACT Objective: To analyze the clinical, laboratory, and radiological findings and management of patients with clinical pituitary apoplexy and to screen for aryl hydrocarbon receptor-interacting protein (AIP) mutations. Subjects and methods: The clinical findings were collected from the medical records of consecutive sporadic pituitary adenoma patients with clinical apoplexy. Possible precipitating factors, laboratory data, magnetic resonance imaging (MRI) findings and treatment were also analyzed. Peripheral blood samples were obtained for DNA extraction from leukocytes, and the entire AIP coding region was sequenced. Results: Thirty-five patients with pituitary adenoma were included, and 23 (67%) had non-functioning pituitary adenomas. Headache was observed in 31 (89%) patients. No clear precipitating factor was identified. Hypopituitarism was observed in 14 (40%) patients. MRI from 20 patients was analyzed, and 10 (50%) maintained a hyperintense signal in MRI performed more than three weeks after pituitary apoplexy (PA). Surgery was performed in ten (28%) patients, and 25 (72%) were treated conservatively with good outcomes. No AIP mutation was found in this cohort. Conclusion: Patients with stable neuroophthalmological impairments can be treated conservatively if no significant visual loss is present. Our radiological findings suggest that hematoma absorption lasts more than that observed in other parts of the brain. Additionally, our study suggests no benefits of AIP mutation screening in sporadic patients with apoplexy.

Prognostic and clinicopathological value of zwint expression levels in patients with lung adenocarcinoma: a systematic review and meta-analysis

The current study found that high Zeste White 10 interactor (ZWINT) expression is related to the poor prognosis of patients with a variety of cancers. This study mainly explored the relationship between the expression level of ZWINT and the prognosis of patients with lung adenocarcinoma (LUAD). Briefly, four English databases and two high-throughput sequencing databases were searched and relevant data for meta-analysis were extracted. Pooled mean difference and 95% confidence interval (CI) were used to assess the relationships between clinical features and the expression of ZWINT. Pooled hazard ratio and 95% CI were also used to assess the relationships between clinical features and the expression level of ZWINT. This meta-analysis was registered in PROSPERO (CRD42021249475). A total of 16 high-quality datasets comprising 2,847 LUAD patients were included in this study. Higher ZWINT expression levels were found in patients younger than 65 years, males, and smokers, and were correlated with advanced TNM stages and poor prognosis. Notably, there was no publication bias in this meta-analysis. Overall, our findings indicate that ZWINT is a potential biomarker for poor prognosis and clinicopathological outcomes of patients with LUAD.

Association of serum levels of secreted frizzled-related protein 5 and Wnt member 5a with glomerular filtration rate in patients with type 2 diabetes mellitus and chronic renal disease: a cross-sectional study

ABSTRACT BACKGROUND: Diabetic nephropathy is a common complication of chronic kidney disease (CKD). ­Inflammation in the kidneys is crucial for promoting development and progression of this complication. Wnt member 5a (Wnt5a) and secreted frizzled-related protein 5 (Sfrp5) are proinflammatory proteins associated with insulin resistance and chronic low-grade adipose tissue inflammation. OBJECTIVE: To determine the correlation between serum Sfrp5 and Wnt5a concentrations and glomerular filtration rate in patients with type 2 diabetes mellitus and CKD. DESIGN AND SETTING: Cross-sectional, comparative and observational study in the Department of Endocrinology, Civil Hospital, Culiacán, Sinaloa, Mexico. METHODS: Eighty individuals with chronic kidney disease were recruited. Their serum Sfrp5 and Wnt5a concentrations were quantified using the enzyme-linked immunosorbent assay (ELISA) test. The statistical analysis consisted of the Mann-Whitney U test for independent samples and Spearman correlation, with statistical significance of P < 0.05. RESULTS: Serum Sfrp5 concentration continually increased through the stages of CKD progression, whereas serum Wnt5a concentration presented its highest levels in stage 3 CKD. Negative correlations between estimated glomerular filtration rate (eGFR) and serum concentrations of Sfrp5 (r = -0434, P = 0.001) and Wnt5a (r = -0481, P = 0.001) were found. CONCLUSIONS: There were negative correlations between serum Sfrp5 and Wnt5a concentrations and eGFR at each stage of CKD, with higher levels in female patients. This phenomenon suggests that Sfrp5 and Wnt5a might be involved in development and evolution towards end-stage renal disease.

Spinal protein kinase A and phosphorylated extracellular signal-regulated kinase signaling are involved in the antinociceptive effect of phytohormone abscisic acid in rats / A proteína quinase A da medula espinhal e a sinalização da quinase fosforilada regulada por sinal extracelular estão envolvidas no efeito antinociceptivo do ácido fito-hormônio abscísico em ratos

Abstract Objective: The phytohormone abscisic acid (ABA) as a signaling molecule exists in various types of organisms from early multicellular to animal cells and tissues. It has been demonstrated that ABA has an antinociceptive effect in rodents. The present study was designed to assess the possible role of PKA and phosphorylated ERK (p-ERK) on the antinociceptive effects of intrathecal (i.t.) ABA in male Wistar rats. Methods: The animals were cannulated intrathecally and divided into different experimental groups (n=6‒7): Control (no surgery), vehicle (received ABA vehicle), ABA-treated groups (received ABA in doses of 10 or 20 µg/rat), ABA plus H.89 (PKA inhibitor)-treated group which received the inhibitor 15 min prior to the ABA injection. Tail-flick and hot-plate tests were used as acute nociceptive stimulators to assess ABA analgesic effects. p-ERK was evaluated in the dorsal portion of the spinal cord using immunoblotting. Results: Data showed that a microinjection of ABA (10 and 20 µg/rat, i.t.) significantly increased the nociceptive threshold in tail flick and hot plate tests. The application of PKA inhibitor (H.89, 100 nM/rat) significantly inhibited ABA-induced analgesic effects. Expression of p-ERK was significantly decreased in ABA-injected animals, which were not observed in the ABA+H.89-treated group. Conclusions: Overall, i.t. administration of ABA (10 µg/rat) induced analgesia and p-ERK down-expression likely by involving the PKA-dependent mechanism.

Resumo Objetivo: O ácido fito-hormônio abscísico (ABA) existe como molécula sinalizadora em vários tipos de organismos, de multicelulares a células e tecidos animais. Foi demonstrado que o ABA tem efeito antinociceptivo em roedores. O presente estudo foi desenhado para avaliar o possível papel da PKA e da ERK fosforilada (p-ERK) nos efeitos antinociceptivos do ABA intratecal (i.t.) em ratos Wistar machos. Métodos: Os animais foram canulados por via i.t. e divididos em diferentes grupos experimentais (n=6‒7): controle (sem cirurgia), veículo (veículo ABA recebido), grupos tratados com ABA (recebeu ABA em doses de 10 ou 20 µg/rato), grupo tratado com ABA mais H.89 (inibidor de PKA) que recebeu o inibidor 15 minutos antes da injeção de ABA. Os testes de movimento da cauda e placa quente foram utilizados como estimuladores nociceptivos agudos para avaliar os efeitos analgésicos da ABA. A p-ERK foi avaliada na porção dorsal da medula espinhal por imunotransferência. Resultados: A microinjeção de ABA (10 e 20 µg/rato, i.t.) aumentou significativamente o limiar nociceptivo nos testes de movimento da cauda e placa quente. A aplicação de inibidor de PKA (H.89, 100 nM/rato) inibiu significativamente os efeitos analgésicos induzidos por ABA. A expressão de p-ERK diminuiu significativamente em animais injetados com ABA que não foram observados no grupo tratado com ABA+H.89. Conclusões: No geral, a administração i.t. de ABA (10 µg/rato) induziu a analgesia e expressão negativa de p-ERK provavelmente envolvendo mecanismo dependente de PKA.

La leptina promueve la expresión de Hic-5 y la formación de puntos de actina por la vía dependiente de FAK-Src en células epiteliales mamarias MCF10A / Leptin induced Hic-5 expression and actin puncta formation by the FAK/Src-dependent pathway in MCF10A mammary epithelial cells

Resumen Introducción. La leptina es una hormona secretada por los adipocitos que se ha relacionado con el proceso de la transición de epitelio a mesénquima (Epithelial- Mesenchymal Transition, EMT). Promueve la migración e invasión de las células del epitelio mamario mediante la activación de las cinasas FAK y Src, un complejo regulador de vías de señalización que favorecen la expresión de las proteínas relacionadas con la formación de estructuras proteolíticas implicadas en la invasión y progresión del cáncer. Recientemente, se ha descrito que la sobreexpresión y activación de la proteína Hic-5 durante el mencionado proceso de transición, favorece la formación de los puntos de actina (indicativa de la formación y funcionalidad de los invadopodios), lo cual promueve la degradación local de los componentes de la matriz extracelular y la metástasis del cáncer. Objetivos. Evaluar el papel de las cinasas FAK y Src sobre la expresión y localización subcelular de Hic-5 y la formación de puntos de actina inducida por la leptina en la línea celular MCF10A de epitelio mamario no tumoral. Materiales y métodos. Se utilizaron los inhibidores específicos de la FAK (PF-573228) y la Src (PP2) para evaluar el papel de ambas cinasas en los niveles de expresión y localización subcelular de la proteína Hic-5 mediante Western blot e inmunofluorescencia, así como la formación de puntos de actina mediante la tinción con faloidina-TRITC en células MCF10A estimuladas con leptina. Resultados. La leptina indujo el incremento en la expresión de Hic-5 y la formación de puntos de actina. El tratamiento previo con los inhibidores de las cinasas FAK (PF-573228) y Src (PP2), promovió la disminución en la expresión de Hic-5 y de los puntos de actina en la línea celular MCF10A de epitelio mamario no tumoral. Conclusión. La leptina indujo la expresión y la localización perinuclear de Hic-5 y la formación de puntos de actina mediante un mecanismo dependiente de la actividad de las cinasas FAK y Src en las células MCF10A.

Abstract Introduction: Leptin is a hormone secreted by adipocytes that has been associated with the epithelial-mesenchymal transition (EMT). Additionally, leptin promotes the migration and invasion of mammary epithelial cells through the activation of FAK and Src kinases, which are part of a regulatory complex of signaling pathways that promotes the expression of proteins related to the formation of proteolytic structures involved in the invasion and progression of cancer. Recently, overexpression and activation of Hic-5 during the EMT have been shown to induce the formation of actin puncta; these structures are indicative of the formation and functionality of invadopodia, which promote the local degradation of extracellular matrix components and cancer metastasis. Objective: To evaluate the role of FAK and Src kinases in the expression of Hic-5 during the epithelial-mesenchymal transition induced by leptin in MCF10A cells. Materials and methods: We used specific inhibitors of FAK (PF-573228) and Src (PP2) to evaluate Hic-5 expression and subcellular localization by Western blot and immunofluorescence assays and to investigate the formation of actin puncta by epifluorescence in MCF10A cells stimulated with leptin. Results: Leptin induced an increase in Hic-5 expression and the formation of actin puncta. Pretreatment with inhibitors of FAK (PF-573228) and Src (PP2) promoted a decrease in Hic-5 expression and actin puncta formation in the non-tumorigenic mammary epithelial cell line MCF10A. Conclusion: In MCF10A cells, leptin-induced Hic-5 expression and perinuclear localization, as well as the formation of actin puncta through a mechanism dependent on the kinase activity of FAK and Src.

Pituitary gigantism: a case series from Hospital de San José (Bogotá, Colombia)

ABSTRACT Introduction Gigantism is a rare pediatric disease characterized by increased production of growth hormone (GH) before epiphyseal closure, that manifests clinically as tall stature, musculoskeletal abnormalities, and multiple comorbidities. Materials and methods Case series of 6 male patients with gigantism evaluated at the Endocrinology Service of Hospital de San José (Bogotá, Colombia) between 2010 and 2016. Results All patients had macroadenomas and their mean final height was 2.01 m. The mean age at diagnosis was 16 years, and the most common symptoms were headache (66%) and hyperhidrosis (66%). All patients had acral changes, and one had visual impairment secondary to compression of the optic chiasm. All patients underwent surgery, and 5 (83%) required additional therapy for biochemical control, including radiotherapy (n = 4, 66%), somatostatin analogues (n = 5, 83%), cabergoline (n = 3, 50%), and pegvisomant (n = 2, 33%). Three patients (50%) achieved complete biochemical control, while 2 patients showed IGF-1 normalization with pegvisomant. Two patients were genetically related and presented a mutation in the aryl hydrocarbon receptor-interacting protein (AIP) gene (pathogenic variant, c.504G>A in exon 4, p.Trp168*), fulfilling the diagnostic criteria of familial isolated pituitary adenoma. Conclusions This is the largest case series of patients with gigantism described to date in Colombia. Transsphenoidal surgery was the first-choice procedure, but additional pharmacological therapy was usually required. Mutations in the AIP gene should be considered in familial cases of GH-producing adenomas.

Methylation status of the PPP1R13L promoter region among lung cancer patients and healthy controls. Analytical cross-sectional study

ABSTRACT BACKGROUND: There is evidence that genetic predisposition and epigenetic alteration (e.g. DNA methylation) play major roles in lung cancer. In our genetic epidemiological studies, rs1970764 in oncogene PPP1R13L was most consistently associated with lung cancer risk. Here, we explored the role of PPP1R13L methylation in lung cancer development. DESIGN AND SETTING: Analytical cross-sectional study (45 lung cancer cases and 45 controls), conducted in China. METHODS: We investigated the DNA methylation status of 2,160 cytosine-phosphate-guanine (CpG) sites in the PPP1R13L promoter region using the EpiTYPER assay of the Sequenom MassARRAY platform. RESULTS: In the whole study group, the methylation levels of CpG-6, CpG-9, CpG-20 and CpG-21 were significantly lower and those of CpG-16 were significantly higher in cases than in controls. Among smokers, the methylation levels at five CpG sites (CpG-6, CpG-11, CpG-15, CpG-20 and CpG-21) were statistically significantly lower among cases. Among men, the methylation levels at four CpG sites (CpG-11, CpG-15, CpG-20 and CpG-21) were significantly lower among cases. Regarding smokers, the methylation levels at CpG-7.8 and CpG-21 among cases and at CpG-22 among controls were significantly lower, compared with nonsmokers. The frequency of positivity for methylation was not significantly different between lung cancer cases and controls (68.22% for cases and 71.87% for controls; P = 0.119). CONCLUSION: Our study on a Chinese population suggests that lung cancer patients have aberrant methylation status (hypomethylation tended to be more frequent) in peripheral blood leukocytes at several CpG sites in the PPP1R13L promoter region and that exposure to smoking may influence methylation status.

O plasma rico em fatores de crescimento no tratamento de pacientes com úlceras venosas / Plasma rich in growth factors in the treatment of patients with venous ulcers

Objetivo geral: Analisar as propriedades do plasma rico em fatores de crescimento (PRGF) autólogo, para tratar pacientes com úlceras venosas. Método: Trata-se de um estudo quase- experimental realizado em um Hospital Universitário, cuja coleta de dados ocorreu no período de maio de 2016 a dezembro de 2017. Amostra composta por 18 pacientes com úlcera venosa, que atenderam aos critérios de inclusão. Os participantes compareceram por 12 semanas consecutivas, ao ambulatório para consulta de enfermagem e troca do curativo, com aplicação do PRGF a cada 15 dias. O preparo do PRGF utilizou a técnica de uma centrifugação e o gluconato de cálcio como ativador plaquetário. Realização da contagem de plaquetas e identificação dos fatores de crescimento no PRGF usado no tratamento. Resultados: Amostra homogênea quanto às variáveis socioeconômicas e clínicas, com predomínio de idosos, sexo masculino, ensino fundamental incompleto e renda entre um e dois salários mínimos. Obtenção de um PRGF com contagem baixa de plaquetas, mediana de 122 x 103 mm3 plaquetas, mas com principais fatores de crescimento plaquetários envolvidos na cicatrização (PDGF-AB, PDGF-BB, TGF-ß, VEGF, EGF e FGF2). Houve alta variabilidade na contagem de plaquetas e na concentração de fatores de crescimento, com correlação forte e significativa entre a quantidade de plaquetas e os fatores de crescimento PDGF-AB e PDGF-BB. As médias de cada fator de crescimento foram 1,52 ng/ml para o PDGF-AB, 1,38 ng/ml para o PDGF-BB, 1,40 ng/ml para o TGF-ß, 0,99 ng/ml para VEGF, 1,57 ng/ml para o EGF e 1.04 ng/ml para o FGF2. O tamanho inicial médio das úlceras neste estudo foi de 29,6 ± 23,9 /cm2 e o percentual médio de redução da área após 12 semanas de tratamento foi de 46,2%. O tamanho das úlceras após tratamento era significativamente menor do que o tamanho inicial das úlceras (p-valor do teste de Wilcoxon igual a 0,001). Entretanto, a redução da área das úlceras, no período das 12 semanas, não apresentou correlação com as concentrações dos fatores de crescimento no PRGF. A percentagem de tecido de epitalização apresentou crescimento em todo período do estudo com aumento significativo ao final de 12 tratamento (média = 58,1%) e o tecido desvitalizado apresentou decrescimento durante todo o período do estudo com redução significativa ao final de 12 semanas (média = 11%). Ao final de 12 semanas observou-se uma melhora significativa do tipo de exsudato nas úlceras e uma diminuição significativa da quantidade de exsudato nas úlceras. Conclusão: Neste estudo, foi confirmada a hipótese nula em que a concentração dos fatores de crescimento presentes no PRGF não apresentou relação estatisticamente significativa com a redução da área das úlceras venosas.

Objective: To analyze the properties of autologous plasma rich in growth factors (PRGF), to treat patients with venous ulcers. Method: It is a quasi-experimental clinical study performed in a University Hospital, the data collection occurred from May 2016 to December 2017. Sample of 18 patients with venous ulcer, who met the inclusion criteria. The participants attended weekly for 12 weeks to the outpatient clinic for nursing consultation and dressing change, with application of the plasma rich in growth factors (PRGF) for each 15 days. The PRGF preparation used the single- centrifugation technique and calcium gluconate as a platelet activator. Realization of platelet count and identification of growth factors in PRGF used in treatment. Platelet counting and identification of growth factors in the PRGF used in the treatment. Results: Homogeneous sample regarding socioeconomic and clinical variables, with predominance of the elderly, male gender, incomplete primary education and income between 1 and 2 minimum wages. A PRGF with a low platelet count, median of 122 x 103 mm3 platelets, but with the main platelet growth factors involved in the healing (PDGF-AB, PDGF-BB, TGF-ß, VEGF, EGF and FGF2). There was a high variability in platelet counts and in the concentration of growth factors, with a strong and significant correlation between the amount of platelets and the growth factors PDGF-AB and PDGF-BB. The means of each growth factor were 1.52 ng / ml for PDGF-AB, 1.38 ng / ml for PDGF-BB, 1.40 ng / ml for TGF-ß, 0.99 ng / ml for VEGF, 1.57 ng / ml for EGF and 1.04 ng / ml for FGF2. The initial mean size of the ulcers in this study was 29.6 ± 23.9 / cm2 and the percentage mean reduction in area after 12 weeks of treatment was 46.2%. The ulcer size after treatment was significantly smaller than the initial ulcer size (Wilcoxon p-value = 0.001). However, the area of ulcer reduction over the 12-week period did not correlate with growth factor concentrations in PRGF. The percentage of epitalization tissue increased throughout the study period with a significant increase at the end of 12 treatments (mean = 58.1%) and devitalized tissue decreased throughout the study period with a significant reduction at the end of 12 weeks (mean = 11%). At the end of 12 weeks there was a significant improvement in the type of exudate in the ulcers and a significant decrease in the amount of exudate in the ulcers. Conclusion: In this study, the null hypothesis was confirmed that the concentration of growth factors present in PRGF did not present statistically significant relationship with the reduction of venous ulcers area.

Objetivo general: Analizar las propiedades del plasma rico en factor de crecimiento autólogo (PRGF) para tratar pacientes con úlcera venosa. Método: Estudio cuasi experimental realizado en un hospital universitario. Datos recolectados entre mayo de 2016 y diciembre de 2017. Muestra compuesta por 18 pacientes con úlcera venosa que cumplieron los criterios de inclusión. Los participantes asistieron semanalmente, durante 12 semanas, a la clínica ambulatoria para consulta de enfermería y cambio de apósito, con aplicación de plasma rico en factor de crecimiento (PRGF) en frecuencia bisemanal. La preparación de PRGF utilizó la técnica de centrifugación; se empleó gluconato de calcio como activador plaquetario. Se efectuó recuento de plaquetas e identificación de los factores de crecimiento en el PRGF utilizado en el tratamiento. Resultados: Muestra homogénea en sus variables socioeconómicas y clínicas, con predominio de ancianos, varones, estudios primarios incompletos, con ingresos de entre 1 y 2 salarios mínimos. Se obtuvo un PRGF con bajo recuento de plaquetas, con mediana de 122 x 103 mm3 de plaquetas, aunque con importantes factores de crecimiento plaquetario involucrados en la curación (PDGF-AB, PDGF-BB, TGF-ß, VEGF, EGF y FGF2). Existió marcada variabilidad en el recuento plaquetario y en la concentración del factor de crecimiento, con marcada y significativa correlación entre la cantidad de plaquetas y factores de crecimiento PDGF-AB y PDGF-BB. Los promedios de cada factor de crecimiento fueron 1.52 ng / ml para PDGF-AB, 1.38 ng / ml para PDGF-BB, 1.40 ng / ml para TGF-ß, 0.99 ng / ml para VEGF, 1,57 ng / ml para EGF y 1,04 ng / ml para FGF2. El tamaño inicial promedio de las úlceras observadas en este estudio fue de 29,6 ± 23,9 / cm2, y la reducción porcentual media en el área luego de 12 semanas de tratamiento fue del 46,2%. El tamaño de las úlceras luego del tratamiento resultó ser significativamente inferior a su tamaño inicial (p-valor del test de Wilcoxon igual a 0,001). Sin embargo, la reducción del área de las úlceras, durante el período de 12 semanas, no se correlacionó con las concentraciones de los factores de crecimiento en el PRGF. El porcentaje de tejido epitelial aumentó durante todo el período del estudio, con un aumento significativo final en 12 tratamientos (media = 58,1%). El tejido desvitalizado disminuyó a lo largo del estudio, con reducción significativa luego de 12 semanas (promedio = 11%). Finalizadas las 12 semanas, hubo mejora significativa en el tipo de exudado de las úlceras y disminución significativa de la cantidad de exudado de las úlceras. Conclusión: En este estudio, se confirmó la hipótesis nula en que la concentración de los factores de crecimiento presentes en el PRGF no presentaba una relación estadísticamente significativa con la reducción del área de las úlceras venosas.

Downregulation of lncRNA CCDC26 contributes to imatinib resistance in human gastrointestinal stromal tumors through IGF-1R upregulation

Imatinib is the first line of therapy for patients with metastatic or gastrointestinal stromal tumors (GIST). However, drug resistance limits the long-term effect of imatinib. Long non-coding RNAs (lncRNAs) are emerging as key players in regulating drug resistance in cancer. In this study, we investigated the association between lncRNA CCDC26 and IGF-1R in GIST and their involvement in drug resistance. Considering the key role of lncRNAs in drug resistance in cancer, we hypothesized that IGF-1R is regulated by lncRNAs. The expression of a series of reported drug resistance-related lncRNAs, including CCDC26, ARF, H19, NBR2, NEAT1, and HOTAIR, in GIST cells treated with imatinib H19 was examined at various time-points by qRT-PCR. Based on our results and published literature, CCDC26, a strongly down-regulated lncRNA following imatinib treatment, was chosen as our research target. GIST cells with high expression of CCDC26 were sensitive to imatinib treatment while knockdown of CCDC26 significantly increased the resistance to imatinib. Furthermore, we found that CCDC26 interacted with c-KIT by RNA pull down, and that CCDC26 knockdown up-regulated the expression of IGF-1R. Moreover, IGF-1R inhibition reversed CCDC26 knockdown-mediated imatinib resistance in GIST. These results indicated that treatments targeting CCDC26-IGF-1R axis would be useful in increasing sensitivity to imatinib in GIST.

Pretentious genomic selection signatures in CYP19A1 gene associated with silent estrous behavior in water buffalo in Pakistan

Background: Poor reproductive efficiency of river buffalos hampers the production capabilities of animals. Buffalos are mainly considered poor breeders owing to the constrained expression of estrus behavior. Failure to display heat signs is an indication of improper functionality of signaling peptides to trigger on a series of behavioral changes, which can be detectable by breeders for timely insemination of females. This might cause an animal to be a repeat breeder. Genomic variations underlying synthesis of signaling peptides can be a useful marker to select superior animals with better reproductive efficiency. In this context, the current study was designed to analyze the CYP19A1 gene in Nili-Ravi buffalo. Results: A total of 97 animals were selected and were divided into two groups on the basis of their heat score. PCR amplification and sequencing of the amplicons were performed using the specific sets of primer, and then, sequences were analyzed for novel variants. A total of 11 polymorphic sites were identified illustrating phenotypic variation in the heat score. Most of the loci were found homologous. Single Nucleotide Polymorphisms (SNPs) were analyzed for association with silent estrus. A three-dimensional protein model was also generated to locate the position of exonic SNPs. Conclusion: This study illustrated that polymorphic sites in the CYP19A1 gene provided potential markers for selection of buffalos with better estrus behavior.

NCS-1 en la función neuronal / NCS-1 in neuronal function

NCS-1 es una proteína unidora de calcio, que regula el funcionamiento de otras proteínas, con las cuales interactúa a nivel molecular. Su expresión es amplia y no limitada a neuronas. Sus efectos incluyen la regulación de receptores, canales iónicos y enzimas que intervienen en múltiples funciones neuronales. NCS-1 regula la actividad del receptor D2 de dopamina y del receptor A2A de adenosina, ambos fundamentales en diversos procesos de comunicación que involucran control emocional y control de movimientos en varios circuitos. NCS-1 también regula la actividad del receptor de IP3, un canal de calcio intracelular fundamental en la regulación de la homeostasis de este ion, interactúa con IP kinasas, las cuales a su vez desencadenan cascadas de señalización intracelular y modula la actividad de canales de calcio presinápticos; todos estos efectos redundan en regulación de la liberación de neurotransmisores y por ende, de la plasticidad sináptica, lo cual ha sido probado en diversos modelos experimentales. NCS-1 también parece estar involucrada en la regulación de otros canales iónicos de calcio y de potasio que podrían influir en la homeostasis eléctrica de las neuronas y en la supervivencia neuronal a través de la regulación de vías proapoptóticas. Estos amplios efectos de NCS-1 motivan a profundizar la investigación en los mecanismos involucrados en la regulación que ejerce sobre sus proteínas blanco y en nuevos efectos que ayuden a entender el rol de esta proteína en diversos procesos fisiológicos y fisiopatológicos(AU)

NCS-1 is a calcium-binding protein, which regulates the functioning of diverse proteins, with which interacts to a molecular level. Its expression is widespread and it is not limited to neurons. Its effects include the regulation of receptors, ion channels and enzymes, which intervene in multiple neuronal functions. NCS-1 regulates the functioning of D2 dopamine receptor and adenosine A2A receptor, both fundamental in diverse communication processes that involve emotional and movement control in a variety of neural circuits. NCS-1 also regulates the activity of IP 3 receptor, an intracellular calcium ion channel (which is crucial in the regulation of calcium homeostasis), interacts with the IP kinases, which trigger intracellular signaling cascades, and modulates the activity of presynaptic calcium channels. All these effects lead to the regulation of neurotransmitters release and thus, synaptic plasticity, which had been proved in diverse experimental models. NCS-1 also appears to be involved in the regulation of other calcium and potassium channels, which can influence the neuron electric homeostasis and survival through the modulation of proapoptotic pathways. These broad NCS-1 effects motivates further research of the specific mechanisms that are involved in the regulation that this protein exerts on its target proteins and in new effects that may help to understand the role of this protein in physiological and pathophysiological processes(AU)

Efficacy of phosphocreatine pre-administration on XIAP and Smac in ischemic penumbra of rats with focal cerebral ischemia reperfusion injury

Abstract Purpose: To observe the efficacy of phosphocreatine pre-administration (PCr-PA) on X-linked inhibitor of apoptosis protein (XIAP), the second mitochondia-derived activator of caspase (Smac) and apoptosis in the ischemic penumbra of rats with focal cerebral ischemia-reperfusion injury (CIRI). Methods: A total of 60 healthy male Sprague Dawley (SD) rats were randomly divided into three groups (n=20): group A (the sham operation group), group B <intraperitoneally injected with 20 mg/kg (10 mg/ml) of saline before preparing the ischemia-reperfusion (IR) model>, and group C <intraperitoneally injected with 20 mg/kg (10 mg/ml) of PCr immediately before preparing the IR model>. After 24 h for reperfusion, the neurological function was evaluated and the tissue was sampled to detect expression of XIAP, Smac and caspase-3 positive cells in the ischemic penumbra so as to observe the apoptosis. Results: Compared with group B, neurological deficit scores, numbers of apoptotic cells, expression of Smac,caspase-9 and the numbers of Caspase-3 positive cells were decreased while expression of XIAP were increased in the ischemic penumbra of group C. Conclusions: Phosphocreatine pre-administration may elicit neuroprotective effects in the brain by increasing expression of X-linked inhibitor of apoptosis protein, reducing expression of second mitochondia-derived activator of caspase, and inhibiting the apoptosis in the ischemic penumbra.

CMPD1 inhibited human gastric cancer cell proliferation by inducing apoptosis and G2/M cell cycle arrest

BACKGROUND: Gastric cancer occupies the fourth highest morbidity rate of cancers worldwide. Clinical therapies of gastric cancer remain limited because of uncertainty of mechanisms and shortness of effective medicine. Thus, new drug candidates for gastric cancer treatment is urgently needed. RESULTS: In this study, CMPD1 as a wildly used MK2 phosphorylation inhibitor was employed to find its impact on gastric cancer cell proliferation, apoptosis and cell cycle using colony formation assay and flow cytometry analysis. Along with its anti-proliferation effect on gastric cancer cell line MKN-45 and SGC7901, CMPD1 also induced massive apoptosis and significant G2/M phase arrest in a time-dependent and dose-dependent manner in MKN-45 cells respectively. Furthermore, Western blot confirmed that the expression of anti-apoptotic proteins Bcl-2 was decreased while BAX, cytochrome c release and cleaved PARP were increased. In addition, oncogene c-Myc was downregulated in response to CMPD1 treatment. CONCLUSIONS: Our results demonstrated that CMPD1 has anti-tumor effect on human gastric cancer cell line MKN- 45 possibly via downregulating oncogene c-Myc expression and CMPD1 could be applied as a potential candidate for treating gastric malignancy. To the best of our knowledge, it is the first report of anti-tumor effect of CMPD-1 on human gastric cancer cells.

Study of major genetic factors involved in pituitary tumorigenesis and their impact on clinical and biological characteristics of sporadic somatotropinomas and non-functioning pituitary adenomas

Genetic and functional aberrations of guanine nucleotide-binding protein, alpha stimulating (GNAS), aryl hydrocarbon receptor interacting protein (AIP), and pituitary tumor transforming gene (PTTG) are among the most prominent events in pituitary tumorigenesis. A cohort of Brazilian patients with somatotropinomas (n=41) and non-functioning pituitary adenomas (NFPA, n=21) from a single tertiary-referral center were evaluated for GNAS and AIP mutations and gene expression of AIP and PTTG. Results were compared to the clinical and biological (Ki67 and p53 expression) characteristics of tumors and their response to therapy, if applicable. Genetic analysis revealed that 27% of somatotropinomas and 4.8% of NFPA harbored GNAS mutations (P=0.05). However, no differences were observed in clinical characteristics, tumor extension, response to somatostatin analog therapy, hormonal/surgical remission rates, Ki67 index, and p53 expression between mutated and non-mutated somatotropinomas patients. PTTG overexpression (RQ mean=10.6, min=4.39, max=11.9) and AIP underexpression (RQ mean=0.56, min=0.46-max=0.92) were found in virtually all cases without a statistically significant relationship with clinical and biological tumor features. No patients exhibited somatic or germline pathogenic AIP mutations. In conclusion, mutations in GNAS and abnormal PTTG and AIP expression had no impact on tumor features and treatment outcomes in this cohort. Our data support some previous studies and point to the need for further investigations, probably involving epigenetic and transcriptome analysis, to improve our understanding of pituitary tumor behavior.

Síndrome de Sotos diagnosticado por hibridación genómica comparativa / Sotos syndrome diagnosed by comparative genomic hybridisation

El síndrome de Sotos (SS) es una enfermedad genética con un patrón de herencia autosómico dominante, causado por haploinsuficiencia del gen NSD1 secundaria a mutaciones puntuales o microdeleciones del locus 5q35 en el que está ubicado el gen. Es un síndrome poco frecuente, presentándose en 7 de cada 100.000 nacimientos. El objetivo de este reporte es presentar el caso de una paciente de 4 años con retardo global del desarrollo, y hallazgos físicos especiales que sugerían un sindrome genético. Caso clínico: Paciente de 4 años, género femenino, cabello ralo, fascie triangular, fisura palpebral alargada, papadar ojival, mandíbula prominente, escápula alada y clinodactilia del quinto dedo de ambas manos. La prueba molecular de hibridación genómica comparativa por microarreglos, mostró microdeleción de la región 5q35.2 q35.3 de 2.082 MB, que incluye el gen NSD1. Conclusión: Proponemos realizar la prueba de hibridación genómica comparativa en pacientes con retraso global del desarrollo y hallazgos fenotípicos menores.

Sotos Syndrome (SS) is a genetic disease with an autosomal dominant pattern caused by haplo-insufficiency of NSD1 gene secondary to point mutations or microdeletion of the 5q35 locus where the gene is located. It is a rare syndrome, occurring in 7 out of every 100,000 births. The objective of this report is to present the case of a 4 year-old patient with a global developmental delay, as well as specific physical findings suggesting a syndrome of genetic origin. Clinical case: Female patient, 4 years of age, thinning hair, triangular facie, long palpebral fissure, arched palate, prominent jaw, winged scapula and clinodactilia of the fifth finger both hands. The molecular test comparative genomic hybridisation test by microarray was subsequently performed, with the result showing 5q35.2 q35.3 region microdeletion of 2,082 MB, including the NSD1 gene. Conclusion: Finally, this article also proposes the performing of comparative genomic hybridisation as the first diagnostic option in cases where clinical findings are suggestive of SS.

Estrogen signaling in the proliferative endometrium: implications in endometriosis / Sinalização pelo estrogênio no endométrio proliferativo: implicações na endometriose

SUMMARY Even though the physiological role of estrogen in the female reproductive cycle and endometrial proliferative phase is well established, the signaling pathways by which estrogen exerts its action in the endometrial tissue are still little known. In this regard, advancements in cell culture techniques and maintenance of endometrial cells in cultures enabled the discovery of new signaling mechanisms activated by estrogen in the normal endometrium and in endometriosis. This review aims to present the recent findings in the genomic and non-genomic estrogen signaling pathways in the proliferative human endometrium specifically associated with the pathogenesis and development of endometriosis.

RESUMO Embora esteja bem estabelecido o papel fisiológico do estrogênio no ciclo reprodutivo feminino e na fase proliferativa do endométrio, as vias de sinalização por meio das quais a ação do estrogênio é exercida no tecido endometrial são ainda pouco conhecidas. Nesse sentido, o avanço nas técnicas de cultura celular e a manutenção de células endometriais em cultivo possibilitaram a descoberta de novos mecanismos sinalizadores ativados pelo estrogênio no endométrio normal e na endometriose. Esta revisão tem o objetivo de apresentar as descobertas recentes envolvendo as vias de sinalização genômica e não genômica do estrogênio no endométrio proliferativo humano, especificamente associadas à patogênese e ao desenvolvimento da endometriose.

Identificación de variantes del gen NPHS2 en niños con síndrome nefrótico corticorresistente / NPHS2 Mutation analysis study in children with steroid-resistant nephrotic syndrome

Resumen: La podocina es una proteína localizada en el diafragma de filtración glomerular donde participa en la regulación de la filtración glomerular. Las mutaciones del gen NPHS2, que codifica a la podocina, son la principal causa de síndrome nefrótico corticorresistente (SNCR) autosómico recesivo en niños. Objetivos: Identificar mutaciones de NPHS2 en niños chilenos con SNCR, y establecer la prevalencia de las variantes más frecuentes en un grupo de adultos sanos. Pacientes y método: Análisis mutacional de NPHS2 en 34 niños chilenos con SNCR. Una vez identificadas las dos variantes de NPHS2 de mayor frecuencia, se realizó un screening de estas mutaciones en 223 adultos sanos. El análisis mutacional se realizó por secuenciación directa de los ocho exones codificantes amplificados por reacción de polimerasa en cadena. La secuenciación del DNA se realizó mediante método fluorométrico y las secuencias fueron evaluadas con el software SeqPilot. La asociación entre la presencia de variantes de NPHS2 y SNCR se calculó comparando las frecuencias alélicas entre los pacientes con SNCR y los voluntarios sanos utilizando prueba exacta de Fisher. Se consideró significativo p < 0,05. Resultados: Se detectaron mutaciones patogénicas de NPHS2 en siete de los 34 pacientes (21%) estudiados, de los cuales seis resultaron heterocigotos para p.R229Q y p.A284 V. En voluntarios sanos la prevalencia de p.R229Q fue de 2,46%. Conclusiones: Este estudio muestra que p.R229Q y p.A284 V son las variantes de NPHS2 más frecuentes en niños chilenos con SNCR. Por primera vez se describe esta asociación en niños chilenos, en base a la cual es posible proponer una estrategia de screening para estudio genético en pacientes con SNCR y sus familias. Se propone una estrategia de búsqueda de p.R229Q y p.A284 V en forma paralela o secuencial en estos pacientes.

Abstract: Podocin is a protein located in the glomerular slit diaphragm where it takes part in the regulation of glomerular filtration. Mutations of the NPHS2 gene that codes podocin are the main cause of autosomal recessive steroid resistant nephrotic syndrome (SRNS). Objectives: To identify the NPHS2 mutations in Chilean children with SRNS, and to determine the prevalence of the most common variants in a group of healthy adults. Patients and methods: Mutation analysis of NPHS2 in 34 Chilean children with SRNS. Once the two most common variants of NPHS2 were identified, screening for these mutations was performed on 233 healthy adults. The mutation analysis was performed by the direct sequencing of the eight coding exons by polymerase chain reaction amplification. The DNA sequencing was performed using a fluorometric method, and then evaluated with SeqPilot™ software. The relationship between the presence of NPHS2 variants and SRNS was calculated by comparing the allele frequency between patients with SRNS and those of the healthy volunteers using the exact Fisher test. A P < .05 was considered significant. Results: Pathogenic NPHS2 mutations were detected in 7 (21%) of the 34 patients studied, of which 6 were heterozygotes for p.R229Q and p.A284 V. The presence of p.R229Q was 2.46% in the healthy volunteers. Conclusions: This study shows that p.R229Q and p.A284 V are the most frequent variants in Chilean children with SRNS. It is the first time that this relationship has been reported in Chilean children. Based on this, a screening strategy is proposed for the genetic study in patients with SRNS and their families. A parallel or sequential search strategy for p.R229Q and p.A284 V in these patients is proposed.

Cuidados em saúde entre portadores de diabetes mellitus autorreferido no Brasil, Pesquisa Nacional de Saúde, 2013 / Health care among adults with self-reported diabetes mellitus in Brazil, National Health Survey, 2013

Resumo: Objetivo: Descrever medidas do cuidado assistencial destinadas ao paciente com diabetes mellitus autorreferido no Brasil. Métodos: Foram utilizados dados da Pesquisa Nacional de Saúde(2013), estudo transversal de base populacional, referentes ao cuidado em saúde com o diabetes mellitus autorreferido, quanto ao uso de serviços de saúde e acesso a medicamentos. Resultados: A prevalência de diabetes mellitus autorreferido foi de 6,2%, e 11,5% da população nunca fez uma glicemia na vida. Dos adultos que referiram diabetes mellitus , 80,2% tomaram medicamentos nas duas semanas anteriores à entrevista, 57,4% usaram o Programa Farmácia Popular, 73,2% receberam assistência médica e 47,1% realizaram o atendimento nas Unidades Básicas de Saúde. Em 65,2%, o médico que atendeu na última consulta era o mesmo das consultas anteriores, 95,3% dos pacientes conseguiram realizar os exames complementares solicitados e 83,3% conseguiram fazer as consultas com o médico especialista. A avaliação de pés e olhos foi relatada por 35,6 e 29,1% dos portadores de diabetes mellitus , respectivamente. Relataram internação hospitalar por causa do diabetes ou de alguma complicação 13,4% dos adultos, e outros 7,0% relataram limitações nas atividades diárias. Em geral, mulheres, assim como a população mais idosa, de maior escolaridade, brancos e residentes nas regiões Sul e Sudeste, tiveram maior prevalência da doença e maior acesso aos serviços, medicamentos e consultas. Discussão: Os cuidados aos portadores de diabetes foram recebidos de forma adequada, na maioria dos casos, o que é essencial para manter a qualidade de vida dos pacientes e prevenir desfechos mais graves.

Abstract: Objective: To describe the care measurements provided to patients with self-reported diabetes mellitus in Brazil. Methods: Data from the Brazilian National Health Survey (2013) were used. This is a cross-sectional population-based study in which the subjects with self-reported diabetes mellitus answered questions concerning their use of health services and access to medicine. Results: The prevalence of self-reported diabetes mellitus was 6.2%, while 11.5% of the population had never undergone a glucose testing. From the adults with diabetes mellitus, 80.2% had taken medications two weeks before the interview, 57.4% used the Popular Pharmacy Program, 73.2% received medical care, and 47.1% were cared for in the Health Basic Units. In 65.2%, the physician who cared for them in the last appointment was the same from previous ones, 95.3% of the patients were able to perform the required complementary examinations, and 83.3% could go to the appointments with a specialist. About 35.6 and 29.1% of the subjects with diabetes mellitus reported feet and eyes examination, respectively. About 13.4% declared previous hospitalization owing to diabetes or any complications, and 7.0% mentioned limitations in their daily activities owing to the disease. In general, women and the elderly people, those with higher education levels, white, and those living in the south and southeastern regions showed a higher prevalence of the disease and greater access to services, medicine, and appointments. Discussion: The care reported by patients with diabetes, which is essential to maintain their quality of life and prevent serious outcomes, seemed, in most cases, to be adequate.