Therapeutic ultrasound stimulates MC3T3-E1 cell proliferation through the activation of NF-κB1, p38α, and mTOR.

BACKGROUND AND OBJECTIVES: As the population ages, osteometabolic diseases and osteoporotic fractures emerge, resulting in substantial healthcare resource utilization and impaired quality of life. Many types of mechanical stimulation have the potential of being recognized by bone cells after a mechanical sign is transformed into a biological one (a process called mechanotransduction). The therapeutic ultrasound (TU) is one of several resources capable of promoting bone cell mechanical stimulation. Therefore, the main purpose of present study was to evaluate the effect of TU on the proliferation of pre-osteoblasts using in vitro bioassays. STUDY DESIGN/MATERIALS AND METHODS: We used MC3T3-E1 pre-osteoblast lineage cells kept in Alpha medium. Cells were treated using pulsed mode therapeutic ultrasound, with frequency of 1 MHz, intensity of 0.2 W/cm(2) (SATA), duty cycle of 20%, for 30 minutes. Nifedipine and rapamycin were used to further investigate the role of L-type Ca(2+) channels and mTOR pathway. Intracellular calcium, TGF-ß1, magnesium, and the mRNA levels of osteopontin, osteonectin, NF-κB1, p38α were evaluated. RESULTS: The results show that TU stimulates the growth of MC3T3-E1 cells and decreases the supernatant calcium and magnesium content. Also, it increases intracellular calcium, activates NF-κB1 and mTOR complex via p38α. Moreover, TU promoted a decrease in the TGF-ß1 synthesis, which is a cell growth inhibitor. CONCLUSIONS: Therapeutic ultrasound, with frequency of 1 MHz, intensity of 0.2 W/cm(2) (SATA) and pulsed mode, for 30 minutes, was able to increase the proliferation of preosteoblast-like bone cells. This effect was mediated by a calcium influx, with a consequent activation of the mTOR pathway, through increased NF-κB1 and p38α.

Characterization of technological and probiotic properties of indigenous Lactobacillus spp. from south Brazil.

In this study, we isolated Lactobacillus spp. from bovine raw milk and artisanal cheese from southern Brazil, and evaluated their technological and probiotic potential to select new isolates for producing healthy fermented dairy foods with differentiated tastes and flavours. We obtained 48 new lactobacilli isolates, which were isolated from raw milk (38) and cheese (10). These bacterial isolates were closely related with ten species: Lactobacillus paracasei (50% of the isolates), L. parabuchneri (15%), L. pentosus (13%), L. zeae (4%), L. plantarum (4%), L. otakiensis (4%), L. casei (4%), L. harbinensis (2%), L. diolivorans (2%), and L. rhamnosus (2%). Isolates CH112 and CH131 showed the greatest acidification potential, reducing the pH of milk to below 5.3 after incubation for 6 h at 32 °C. Considering proteolytic activity and diacetyl production, isolates ML88a, ML04, and ML12 showed the most promising results. Isolate ML12 showed 100% survival rate when inoculated in gastric juice at pH 2.5. The evaluation of antibacterial activity of the lactobacilli showed that the pathogens Listeria monocytogenes, Staphylococcus aureus, Salmonella enteritidis, and Salmonella Typhimurium were strongly inhibited by the pure lactobacilli cultures. Five Lactobacillus isolates (ML01, ML04, ML12, ML88, and CH139) showed both technological and probiotic characteristics. Principal Component Analysis (PCA) was used to investigate correlations among technological and probiotic characteristics, and identified new promising lactobacilli isolates for exploring their characteristics. This study reveals the importance of selecting new microorganisms with potential applicability in the food industry for developing functional foods with differentiated aromas and flavours.

Calyptranthes grandifolia O.Berg (Myrtaceae) ethanolic extract inhibits TNF-α gene expression and cytokine release in vitro.

Anti-tumor therapies based on anti-inflammatory effects have been considered in cancer treatment. Survival, proliferation and, resultantly, invasion and metastasis of tumor cells are regulated by local inflammatory mediators. Primary inflammatory cytokines, such as tumor necrosis factor (TNF), are targets for anticancer therapy. Several anti­inflammatory agents isolated from natural products are becoming important chemopreventive and therapeutic agents for cancer. The present study aimed to investigate the expression of TNF­α, nuclear factor­κΒ (NF­κΒ) and p38α mitogen-activated protein kinase (p38α) genes, associated with proliferation and inflammation in the Caco­2 cell line treated with ethanolic and hexanic extracts of Calyptranthes grandifolia O.Berg (Myrtaceae). Caco­2 cells were cultured and treated with plant extract at different concentrations (25, 50, 100 and 200 µg/ml) and stimulated with lipopolysaccharide (LPS). For gene expression, analysis was performed by total RNA extraction followed by synthesis of complementary DNA and analysis by quantitative polymerase chain reaction. The release of TNF­α cytokine was evaluated by ELISA in RAW 264.7 murine macrophages activated by LPS. Among the evaluated genes, there was a decrease in TNF-α expression at 100 and 200 µg/ml concentrations only with the ethanolic extract (P<0.025). The p38α gene exhibited a tendency to increase expression only when treated with ethanolic extract and the NF­κΒ gene did not significantly differ compared with the positive control when treated with either analyzed extract. The inhibition of TNF-α cytokine in the RAW 264.7 cell line was significant (P<0.05) in ethanolic extract at 200 µg/ml compared with the positive control (LPS 1 µg/ml). In conclusion, the ethanolic extract may exhibit an anti­inflammatory activity by inhibiting TNF­α. However, further studies are required to confirm its potential anti-inflammatory effects.

Effect of Helicobacter pylori on NFKB1, p38α and TNF-α mRNA expression levels in human gastric mucosa.

Helicobacter pylori infects ~50% of the world population, causing chronic gastritis and other forms of cellular damage. The present study assessed the influence of H. pylori on the mRNA expression levels of nuclear factor-κB1 (NFKB1), p38α and tumor necrosis factor-α (TNF-α) in human gastric mucosa in a southern Brazilian population. Human gastric tissue was collected by upper endoscopy and H. pylori diagnosis was performed using a rapid urease test and histological analysis. Total RNA was extracted and purified for subsequent cDNA synthesis and analysis by quantitative polymerase chain reaction (qPCR). The gastric tissue samples were divided into four groups as follows: Normal, inactive chronic gastritis, active chronic gastritis and intestinal metaplasia. The SDHA gene was classified as the most stable when compared with ACTB, GAPDH, B2M and HPRT1 genes, and was therefore selected as the reference gene for qPCR data normalization. TNF-α mRNA expression was significantly higher in samples that were positive for H. pylori and with active chronic gastritis. However, no difference was detected in the mRNA expression levels of NFKB1 and p38α between the groups. The present study concluded that the presence of H. pylori is associated with TNF-α upregulation in human gastric mucosa, but had no effect on NFKB1 and p38α mRNA expression levels.

Association study of C936T polymorphism of the VEGF gene and the C242T polymorphism of the p22phox gene with diabetes mellitus type 2 and distal diabetic polyneuropathy.

Even with long­term glycemic control, diabetes mellitus type 2 (DM2) remains the predominant cause of diabetic neuropathy. Single nucleotide polymorphism (SNP) C936T of the vascular endothelial growth factor (VEGF) gene and the SNP C242T of the p22phox (CYBA) gene have been investigated in relation to DM2 and its complications. The aim of the present study was to investigate the association between these two SNPs and DM2, and also between the SNPs and the signs and symptoms of diabetic distal polyneuropathy. The DM2 group consisted of 98 individuals and the control group consisted of 104 individuals. The results demonstrated that there was no association between the different genotypes or alleles and increased risk of the disease (P>0.05). With SNP C242T, a significant association with body mass index between the CTxTT genotypes (P=0.043) was identified; and the greatest body mass indexes were among individuals with the TT genotype. An association between the degree of neuropathic symptoms and genotypic/allelic distribution of these polymorphisms was not observed. In conclusion, the investigated polymorphisms are not correlated with the risk of developing DM2.

Celiac plexus block: an anatomical study and simulation using computed tomography.

OBJECTIVE: To analyze anatomical variations associated with celiac plexus complex by means of computed tomography simulation, assessing the risk for organ injury as the transcrural technique is utilized. MATERIALS AND METHODS: One hundred eight transaxial computed tomography images of abdomen were analyzed. The aortic-vertebral, celiac trunk (CeT)-vertebral, CeT-aortic and celiac-aortic-vertebral topographical relationships were recorded. Two needle insertion pathways were drawn on each of the images, at right and left, 9 cm and 4.5 cm away from the midline. Transfixed vital organs and gender-related associations were recorded. RESULTS: Aortic-vertebral - 45.37% at left and 54.62% in the middle; CeT-vertebral - T12, 36.11%; T12-L1, 32.4%; L1, 27.77%; T11-T12, 2.77%; CeT-aortic - 53.7% at left and 46.3% in the middle; celiac-aortic-vertebral - L-l, 22.22%; M-m, 23.15%; L-m, 31.48%; M-l, 23.15%. Neither correspondence on the right side nor significant gender-related associations were observed. CONCLUSION: Considering the wide range of abdominal anatomical variations and the characteristics of needle insertion pathways, celiac plexus block should not be standardized. Imaging should be performed prior to the procedure in order to reduce the risks for injuries or for negative outcomes to patients. Gender-related anatomical variations involved in celiac plexus block should be more deeply investigated, since few studies have addressed the subject.

OBJETIVO: Analisar variações anatômicas relacionadas ao bloqueio do plexo celíaco por meio da simulação por tomografia computadorizada e avaliar a possibilidade de transfixação de órgãos pelo método transcrural. MATERIAIS E MÉTODOS: Cento e oito imagens de tomografias computadorizadas transaxiais abdominais foram analisadas. As relações aorto-vertebral, tronco celíaco (TCe)-vertebral, TCe-aórtica e celíaco-aorto-vertebral foram registradas. Em cada imagem foram dispostas duas trajetórias de agulhas, a 9 cm e a 4,5 cm à esquerda e à direita da linha média. Os órgãos vitais transfixados e associações relacionadas ao gênero foram registrados. RESULTADOS: Aorto-vertebral - 45,37% esquerda e 54,62% central; TCe-vertebral - T12, 36,11%; T12-L1, 32,4%; L1, 27,77%; T11-T12, 2,77%; TCe-aórtica - 53,7% esquerda e 46,3% central; celíaco-aorto-vertebral - L-l, 22,22%; M-m, 23,15%; L-m, 31,48%; M-l, 23,15%. Em nenhum dos critérios analisados houve correspondência no lado direito e nem associação significativa entre os gêneros. CONCLUSÃO: O bloqueio do plexo celíaco não deve ser padronizado, em razão das amplas variações anatômicas abdominais e das características próprias de cada acesso, sendo necessário o registro de imagem prévio ao procedimento para cada paciente, visando diminuir riscos de lesão. Registros sobre a variação anatômica quanto ao gênero, relacionados ao bloqueio do plexo celíaco, devem ser aprofundados.

Verificação do dano muscular através da avaliação da concentração decreatina quinase em indivíduos não atletas após prova de ciclismo não competitivo / Evaluation of muscle damage by assessment of the concentration of creatinekinase in non-athlete individuals after uncompetitive cycling race

Objetivos: Investigar os níveis séricos da enzima creatina quinase em ciclistas não profissionais após uma prova de ciclismo não competitivo de 300 km, para avaliação de dano muscular.Métodos: Participaram do estudo 12 ciclistas do sexo masculino, com as seguintes médias para os parâmetros antropométricos: idade 38,1±11,5 anos, estatura 1,77±7,7 cm, peso 77±8,5 kg, percentual de gordura 12,0±4,1% e índice de massa corporal 24,7±2,6kg/m². Os níveis séricos de creatina quinase foram avaliados antes e imediatamente após a prova, através de método cinético.Resultados: O estudo demonstrou que após a prova de 300 km houve uma elevação média de 480% nos níveis séricos de creatina quinase (p<0,0001). Nenhuma correlação foi encontrada entre os níveis de creatina quinase e o índice de massa corporal ou o tempo de prova.Conclusões: Estes dados demonstram elevação nos níveis séricos de creatina quinase após uma prova de ciclismo não competitivo de 300 km, sugerindo a presença de dano muscular nessa modalidade de ciclismo.

Aims: To investigate the serum levels of the enzyme creatine kinase in non-professional cyclists after a non-competitive cycling race of 300 km, for evaluation of muscle damage.Methods: The study included 12 male cyclists, with the following means for the anthropometric parameters: age 38.1±11.5 years, height 1.77±7.7 cm, weight 77±8.5 kg, fat percentage 12.0±4.1%, and body mass index 24.7±2.6 kg/m². Serum levels of creatine kinase were assessed before and immediately after the race, through kinetic method.Results: The study showed that after the 300 km race there was an average increase of 480% in serum levels of creatine kinase (p<0.0001). No correlation was found between creatine kinase levels and body mass index or time of race.Conclusions: These data show increase in serum levels of creatine kinase after a non-competitive cycling race of 300 km, suggesting the presence of muscle damage in this cycling mode.

Prevalência e perfil sociodemográfico de diabetes e hipertensão em indivíduos do sistema de informação da atenção básica / Prevalence and sociodemographic profile of diabetes and hypertension in individuals registered in the Primary Healthcare Information System / Prevalencia y perfil sociodemográfico de la diabetes e hipertensión de individuos del sistema de información de La atención básica

Objetivos: Verificar a prevalência e o perfil sociodemográfico de indivíduos com diabetes mellitus (DM) e hipertensão arterial sistêmica (HAS), cadastrados no Sistema de Informação da Atenção Básica (SIAB) do município de Lajeado-RS, no ano de 2012. Métodos: O presente estudo, transversal, descritivo-exploratório, com abordagem quantitativa, utilizou dados secundários extraídos do SIAB de 757 indivíduos. Os dados foram processados e analisados com software SPPS versão 20.0. Resultados: Da amostra de 757 indivíduos, obteve-se uma média de idade de 40,87 ± 16,15 anos, sendo 50,2% (380) do sexo masculino e 49,8% (377) do sexo feminino. Da mesma forma, 97% (734) são alfabetizados. Encontrou-se uma frequência de apenas 3,1% (1.131) de DM na população total e de 4,5% (34) na amostra selecionada, e obteve-se uma prevalência de HAS de 15% (5.849) e 17,3% (131), respectivamente. Houve correlação positiva ? fraca, mas significativa ? entre HAS, DM e idade (p=0,000). Conclusão: Os dados obtidos indicaram uma baixa prevalência de DM e HAS em indivíduos cadastrados no SIAB em um município do interior do Rio Grande do Sul.

Objective: To determine the prevalence and sociodemographic profile of individuals with diabetes mellitus (DM) and systemic arterial hypertension (SAH), registered in the Primary Healthcare Information System (Sistema de Informação da Atenção Básica - SIAB) in the city of Lajeado, Rio Grande do Sul, in 2012. Methods: This cross-sectional, descriptive exploratory study, with a quantitative approach, used secondary data retrieved from the SIAB, regarding 757 individuals. Data was processed and analyzed with SPSS software version 20.0. Results: In the sample of 757 subjects, a mean age of 40.87 ± 16.15 years was obtained, with 50.2% (380) male and 49.8% (377) female. Similarly, 97% (734) are literate. The study found a DM frequency of only 3.1% (1,131) in the total population, and of 4.5% (34) in the selected sample, and a prevalence of hypertension of 15% (5,849) and 17.3% (131), respectively. There was a positive, weak but significant, correlation between SAH, DM and age (p=0.000). Conclusion: The data obtained indicates a low prevalence of DM and SAH in individuals registered in SIAB in a countryside city of Rio Grande do Sul.

Objetivo: Verificar la prevalencia y el perfil sociodemográfico de individuos con diabetes mellitus (DM) y hipertensión arterial sistémica (HAS) registrados en el Sistema de Información de La Atención Básica (SIAB) del municipio de Lajeado-RS, en el año de 2012. Métodos: El presente estudio transversal, descriptivoexploratorio con abordaje cuantitativo utilizó datos secundários de 757 individuos extraídos del SIAB. Los datos fueron procesados y analizados en el software SPPS versión 20.0. Resultados: De La muestra de 757 individuos, se obtuvo una media de edad de 40,87 ± 16,15 años, siendo el 50,2% (380) del sexo masculino y el 49,8% (377) del sexo femenino. De la misma manera, el 97% (734) son alfabetizados. Se encontró una frecuencia de solo el 3,1% (1.131) de DM en la población total y del 4,5% (34) en la muestra seleccionada y se obtuvo una prevalencia de HAS del 15% (5.849) y el 17,3% (131), respectivamente. Hubo correlación positiva ? débil, pero significativa ? entre HAS, DM y edad (p=0,000). Conclusión: Los datos obtenidos indicaron baja prevalencia de DM y HAS en individuos registrados en el SIAB de un município del interior de Rio Grande do Sul.

Celiac plexus block: an anatomical study and simulation using computed tomography / Bloqueio do plexo celíaco: estudo anatômico e simulação em tomografia computadorizada

Objective: To analyze anatomical variations associated with celiac plexus complex by means of computed tomography simulation, assessing the risk for organ injury as the transcrural technique is utilized. Materials and Methods: One hundred eight transaxial computed tomography images of abdomen were analyzed. The aortic-vertebral, celiac trunk (CeT)-vertebral, CeT-aortic and celiac-aortic-vertebral topographical relationships were recorded. Two needle insertion pathways were drawn on each of the images, at right and left, 9 cm and 4.5 cm away from the midline. Transfixed vital organs and gender-related associations were recorded. Results: Aortic-vertebral - 45.37% at left and 54.62% in the middle; CeT-vertebral - T12, 36.11%; T12-L1, 32.4%; L1, 27.77%; T11-T12, 2.77%; CeT-aortic - 53.7% at left and 46.3% in the middle; celiac-aortic-vertebral - L-l, 22.22%; M-m, 23.15%; L-m, 31.48%; M-l, 23.15%. Neither correspondence on the right side nor significant gender-related associations were observed. Conclusion: Considering the wide range of abdominal anatomical variations and the characteristics of needle insertion pathways, celiac plexus block should not be standardized. Imaging should be performed prior to the procedure in order to reduce the risks for injuries or for negative outcomes to patients. Gender-related anatomical variations involved in celiac plexus block should be more deeply investigated, since few studies have addressed the subject. .

Objetivo: Analisar variações anatômicas relacionadas ao bloqueio do plexo celíaco por meio da simulação por tomografia computadorizada e avaliar a possibilidade de transfixação de órgãos pelo método transcrural. Materiais e Métodos: Cento e oito imagens de tomografias computadorizadas transaxiais abdominais foram analisadas. As relações aorto-vertebral, tronco celíaco (TCe)-vertebral, TCe-aórtica e celíaco-aorto-vertebral foram registradas. Em cada imagem foram dispostas duas trajetórias de agulhas, a 9 cm e a 4,5 cm à esquerda e à direita da linha média. Os órgãos vitais transfixados e associações relacionadas ao gênero foram registrados. Resultados: Aorto-vertebral - 45,37% esquerda e 54,62% central; TCe-vertebral - T12, 36,11%; T12-L1, 32,4%; L1, 27,77%; T11-T12, 2,77%; TCe-aórtica - 53,7% esquerda e 46,3% central; celíaco-aorto-vertebral - L-l, 22,22%; M-m, 23,15%; L-m, 31,48%; M-l, 23,15%. Em nenhum dos critérios analisados houve correspondência no lado direito e nem associação significativa entre os gêneros. Conclusão: O bloqueio do plexo celíaco não deve ser padronizado, em razão das amplas variações anatômicas abdominais e das características próprias de cada acesso, sendo necessário o registro de imagem prévio ao procedimento para cada paciente, visando diminuir riscos de lesão. Registros sobre a variação anatômica quanto ao gênero, relacionados ao bloqueio do plexo celíaco, devem ser aprofundados. .

Avaliação do perfil glicêmico de pacientes com diabetes mellitus tipo 2 com e sem administração de infusão de folhas de Averrhoa carambola / Evaluation of glycemic profile of patients with diabetes mellitus type 2with or without administration of Averrhoa carambola infusion

Objetivos: avaliar o efeito da infusão das folhas da planta Averrhoa carambola nos níveis glicêmicos de pacientes com diabetes mellitus tipo 2. Métodos: este ensaio clínico estudou 22 pacientes, divididos em grupo teste (11 pacientes que fizeram uso do chá das folhas de Averrhoa carambola) e grupo controle (11 pacientes sem uso do chá), que foram acompanhados durante um período de três meses. Para avaliar os níveis glicêmicos, foi feita dosagem sanguínea em intervalos de 15 dias. Resultados: após três meses, os níveis de glicose sanguínea do grupo que ingeriu o chá das folhas de Averrhoa carambola não diferiram dos níveis do grupo teste. Durante o período da pesquisa a função renal foi monitorada através da dosagem sérica de creatinina, que se mostrou inalterada nos dois grupos. Conclusões: não foram detectados efeitos da ingestão de infusão das folhas da planta Averrhoa carambola nos níveis glicêmicos de pacientes com diabetes mellitus tipo 2.

Aims: To evaluate the effect of Averrhoa carambola infusion in blood glucose levels of patients with Diabetes Mellitus Type 2. Methods: This clinical trial studied 22 patients, divided in test group (11 patients who drank the tea of Averrhoa carambola leaves) and control group (11 patients who did not drink the tea), which were monitored over a period of three months. To analyze the blood glucose levels, blood dosage was done every 15 days. Results: After three months, the blood glucose levels in the group that ingested the tea of leaves of Averrhoa carambola did not differ from those of the test group. During the trial period the renal function was monitored by seric creatinine dosage, which remained unchanged in both groups. Conclusions: This study did not detect effects of the tea of Averrhoa carambola leaves on blood glucose levels of patients with diabetes mellitus Type 2.

Androgen-dependent expression of c-jun and c-fos in human non-transformed epithelial prostatic cells: association with cell proliferation.

OBJECTIVE: To assess the effect of dihydrotestosterone (DHT) on the gene expression of C-FOS and C-JUN and on the proliferation of human non-transformed epithelial prostatic (HNTEP) cells. METHODS: Cell proliferation (MTT) and C-FOS and C-JUN mRNA expression (RT-PCR) were determined in cells treated with DHT (10(-8), 10(-10), and 10(-13)M) or with control medium. RESULTS: DHT 10(-13) M had a significant stimulatory effect on cell proliferation (p < 0.05) and C-FOS and C-JUN gene expression when compared to cells treated with higher concentrations of this hormone (10(-10) and 10(-8)M) or with the control group. CONCLUSIONS: Our data demonstrate that the increase in C-FOS and C-JUN expression and cell growth in HNTEP cells is maximal with the lowest DHT concentration (10(-13)M). These proto-oncogenes may play a role in the control of hormone responsiveness and cell proliferation in HNTEP cells.