Dexamethasone blocks the migration of the human neuroblastoma cell line SK-N-SH

Glucocorticoids (Gc) influence the differentiation of neural crest-derived cells such as those composing sympathoadrenal tumors like pheochromocytomas, as well as neuroblastomas and gangliomas. In order to obtain further information on the effects of Gc on cells evolving from the neural crest, we have used the human neuroblastoma cell line SK-N-SH to analyze: 1) the presence and the binding characteristics of Gc receptors in these cells, 2) the effect of dexamethasone (Dex) on the migration of SK-N-SH cells, and 3) the effect of Dex on the organization of the cytoskeleton of SK-N-SH cells. We show that: 1) receptors that bind [³H]-Dex with high affinity and high capacity (Kd of 9.6 nM, Bmax of 47 fmol/mg cytosolic protein, corresponding to 28,303 sites/cell) are present in cytosolic preparations of SK-N-SH cells, and 2) treatment with Dex (in the range of 10 nM to 1 µM) has an inhibitory effect (from 100% to 74 and 43%, respectively) on the chemotaxis of SK-N-SH cells elicited by fetal bovine serum. This inhibition is completely reversed by the Gc receptor antagonist RU486 (1 µM), and 3) as demonstrated by fluorescent phalloidin-actin detection, the effect of Dex (100 nM) on SK-N-SH cell migration is accompanied by modifications of the cytoskeleton organization that appear with stress fibers. These modifications did not take place in the presence of 1 µM RU486. The present data demonstrate for the first time that Dex affects the migration of neuroblastoma cells as well as their cytoskeleton organization by interacting with specific receptors. These findings provide new insights on the mechanism(s) of action of Gc on cells originating in the neural crest.

Autonomic modulation of heart rate of young and postmenopausal women undergoing estrogen therapy

The aim of the present study was to determine whether estrogen therapy (ET) reduces alterations of the autonomic control of heart rate (HR) due to hypoestrogenism and aging. Thirteen young (24 ± 2.6 years), 10 postmenopausal (53 ± 4.6 years) undergoing ET (PM-ET), and 14 postmenopausal (56 ± 2.6 years) women not undergoing ET (PM) were studied. ET consisted of 0.625 mg/day conjugated equine estrogen. HR was recorded continuously for 8 min at rest in the supine and sitting positions. HR variability (HRV) was analyzed by time (SDNN and rMSSD indices) and frequency domain methods. Power spectral components are reported as normalized units (nu) at low (LF) and high (HF) frequencies, and as LF/HF ratio. Intergroup comparisons: SDNN index was higher in young (median: supine, 47 ms; sitting, 42 ms) than in PM-ET (33; 29 ms) and PM (31; 29 ms) women (P < 0.05). PM showed lower HFnu, higher LFnu and higher LF/HF ratio (supine: 44, 56, 1.29; sitting: 38, 62, 1.60) than the young group in the supine position (61, 39, 0.63) and the PM-ET group in the sitting position (57, 43, 0.75; P < 0.05). Intragroup comparisons: HR was lower in the supine than in the sitting position for all groups (P < 0.05). The HRV decrease from the supine to the sitting position was significant only in the young group. These results suggest that HRV decreases during aging. ET seems to attenuate this process, promoting a reduction in sympathetic activity on the heart and contributing to the cardioprotective effect of estrogen hormones.

Antimicrobial activity of the essential oil from Lippia sidoides, carvacrol and thymol against oral pathogens

Dental caries and periodontal disease are associated with oral pathogens. Several plant derivatives have been evaluated with respect to their antimicrobial effects against such pathogenic microorganisms. Lippia sidoides Cham (Verbenaceae), popularly known as "Alecrim-pimenta" is a typical shrub commonly found in the Northeast of Brazil. Many plant species belonging to the genus Lippia yield very fragrant essential oils of potential economic value which are used by the industry for the commercial production of perfumes, creams, lotions, and deodorants. Since the leaves of L. sidoides are also extensively used in popular medicine for the treatment of skin wounds and cuts, the objective of the present study was to evaluate the composition and antimicrobial activity of L. sidoides essential oil. The essential oil was obtained by hydro-distillation and analyzed by GC-MS. Twelve compounds were characterized, having as major constituents thymol (56.7%) and carvacrol (16.7%). The antimicrobial activity of the oil and the major components was tested against cariogenic bacterial species of the genus Streptococcus as well as Candida albicans using the broth dilution and disk diffusion assays. The essential oil and its major components thymol and carvacrol exhibited potent antimicrobial activity against the organisms tested with minimum inhibitory concentrations ranging from 0.625 to 10.0 mg/mL. The most sensitive microorganisms were C. albicans and Streptococcus mutans. The essential oil of L. sidoides and its major components exert promising antimicrobial effects against oral pathogens and suggest its likely usefulness to combat oral microbial growth.

Cloning, sequence analysis, and expression of cDNA coding for the major house dust mite allergen, Der f 1, in Escherichia coli

Our objective was to clone, express and characterize adult Dermatophagoides farinae group 1 (Der f 1) allergens to further produce recombinant allergens for future clinical applications in order to eliminate side reactions from crude extracts of mites. Based on GenBank data, we designed primers and amplified the cDNA fragment coding for Der f 1 by nested-PCR. After purification and recovery, the cDNA fragment was cloned into the pMD19-T vector. The fragment was then sequenced, subcloned into the plasmid pET28a(+), expressed in Escherichia coli BL21 and identified by Western blotting. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Sequence analysis showed the presence of an open reading frame containing 966 bp that encodes a protein of 321 amino acids. Interestingly, homology analysis showed that the Der p 1 shared more than 87% identity in amino acid sequence with Eur m 1 but only 80% with Der f 1. Furthermore, phylogenetic analyses suggested that D. pteronyssinus was evolutionarily closer to Euroglyphus maynei than to D. farinae, even though D. pteronyssinus and D. farinae belong to the same Dermatophagoides genus. A total of three cysteine peptidase active sites were found in the predicted amino acid sequence, including 127-138 (QGGCGSCWAFSG), 267-277 (NYHAVNIVGYG) and 284-303 (YWIVRNSWDTTWGDSGYGYF). Moreover, secondary structure analysis revealed that Der f 1 contained an a helix (33.96%), an extended strand (17.13%), a ß turn (5.61%), and a random coil (43.30%). A simple three-dimensional model of this protein was constructed using a Swiss-model server. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Alignment and phylogenetic analysis suggests that D. pteronyssinus is evolutionarily more similar to E. maynei than to D. farinae.

Antioxidant and antimicrobial activities of bitter and sweet apricot (Prunus armeniaca L.) kernels

Abstract The present study describes the in vitro antimicrobial and antioxidant activity of methanol and water extracts of sweet and bitter apricot (Prunus armeniaca L.) kernels. The antioxidant properties of apricot kernels were evaluated by determining radical scavenging power, lipid peroxidation inhibition activity and total phenol content measured with a DPPH test, the thiocyanate method and the Folin method, respectively. In contrast to extracts of the bitter kernels, both the water and methanol extracts of sweet kernels have antioxidant potential. The highest percent inhibition of lipid peroxidation (69%) and total phenolic content (7.9 ± 0.2 µg/mL) were detected in the methanol extract of sweet kernels (Hasanbey) and in the water extract of the same cultivar, respectively. The antimicrobial activities of the above extracts were also tested against human pathogenic microorganisms using a disc-diffusion method, and the minimal inhibitory concentration (MIC) values of each active extract were determined. The most effective antibacterial activity was observed in the methanol and water extracts of bitter kernels and in the methanol extract of sweet kernels against the Gram-positive bacteria Staphylococcus aureus. Additionally, the methanol extracts of the bitter kernels were very potent against the Gram-negative bacteria Escherichia coli (0.312 mg/mL MIC value). Significant anti-candida activity was also observed with the methanol extract of bitter apricot kernels against Candida albicans, consisting of a 14 mm in diameter of inhibition zone and a 0.625 mg/mL MIC value.

Correlation between total nitrite/nitrate concentrations and monoamine oxidase (types A and B) and semicarbazide-sensitive amine oxidase enzymatic activities in human mesenteric arteries from non-diabetic and type 2 diabetic patients

The aim of this study was to determine the correlation between total nitrite/nitrate concentrations (NOx) and the kinetic parameters of monoamine oxidase enzymes (MAO-A and MAO-B) and semicarbazide-sensitive amine oxidase (SSAO) in human mesenteric arteries. Arteries were from non-diabetic and type 2 diabetic patients with sigmoid or rectum carcinoma for whom surgery was the first option and who were not exposed to neo-adjuvant therapy. Segments of human inferior mesenteric arteries from non-diabetic (61.1 ± 8.9 years old, 7 males and 5 females, N = 12) and type 2 diabetic patients (65.8 ± 6.2 years old, 8 males and 4 females, N = 12) were used to determine NOx concentrations and the kinetic parameters of MAO-A, MAO-B and SSAO by the Griess reaction and by radiochemical assay, respectively. The NOx concentrations in arteries from diabetic patients did not differ significantly from those of the non-diabetic group (10.28 ± 4.61 vs 10.71 ± 4.32 nmol/mg protein, respectively). In the non-diabetic group, there was a positive correlation between NOx concentrations and MAO-B parameters: Km (r = 0.612, P = 0.034) and Vmax (r = 0.593, P = 0.042), and a negative correlation with the SSAO parameters: Km (r = -0.625, P = 0.029) and Vmax (r = -0.754, P = 0.005). However, in the diabetic group no correlation was found between NOx concentrations and the three kinetic parameters of the enzymes. These results suggest an important function of sympathetic nerves and vascular NOx concentrations in arteries of non-diabetic patients. Thus, these results confirm the importance of a balance between oxidants and antioxidants in the maintenance of vascular homeostasis to prevent oxidative stress.

Clinical features of patients with type 2 diabetes mellitus and hepatitis C infection

The objective of the present cross-sectional study was to assess the prevalence and the clinical and laboratory features of hepatitis C virus (HCV)-positive patients with type 2 diabetes mellitus (DM) attending either an outpatient clinic or hemodialysis units. Serologic-HCV testing was performed in 489 type 2 DM patients (303 outpatients and 186 on dialysis). A structured assessment of clinical, laboratory and DM-related complications was performed and the patients were then compared according to HCV infection status. Mean patient age was 60 years; HCV positivity (HCV+) was observed in 39 of 303 (12.9%) outpatients and in 34 of 186 (18.7%) dialysis patients. Among HCV+ patients, 32 were men (43.8%). HCV+ patients had higher serum levels of aspartate aminotransferase (0.90 ± 0.83 vs 0.35 ± 0.13 µKat/L), alanine aminotransferase (0.88 ± 0.93 vs 0.38 ± 0.19 µKat/L), gamma-glutamyl transferase (1.57 ± 2.52 vs 0.62 ± 0.87 µKat/L; P < 0.001), and serum iron (17.65 ± 6.68 vs 14.96 ± 4.72 µM; P = 0.011), and lower leukocyte and platelet counts (P = 0.010 and P < 0.001, respectively) than HCV-negative (HCV-) patients. HCV+ dialysis patients had higher diastolic blood pressure than HCV- patients (87.5 ± 6.7 vs 81.5 ± 6.0 mmHg; P = 0.005) and a lower prevalence of diabetic retinopathy (75 vs 92.7%; P = 0.007). In conclusion, our study showed that HCV is common among subjects with type 2 DM but is not associated with a higher prevalence of chronic diabetic complications.

Is a contrast study really necessary prior to ureteroscopy?

This study aimed to evaluate the effect of preoperative imaging techniques on the success and complication rates of ureteroscopy. We performed a retrospective analysis of 736 patients (455 males and 281 females), with a mean age of 45.5±15.2 years (range, 1-88 years), who underwent rigid ureteroscopic procedures for removal of ureteral stones. Patients were divided into 4 groups according to the type of imaging modality used: group I, intravenous urography (n=116); group II, computed tomography (n=381); group III, computed tomography and intravenous urography (n=91), and group IV, ultrasonography and abdominal plain film (n=148). Patients’ demographics, stone size and location, prior shock wave lithotripsy, lithotripsy technique, operation time, success rate, and rate of intraoperative complications were compared among the groups. There were no significant differences in success and complication rates among the groups. The stone-free rate after primary ureteroscopy was 87.1% in group I, 88.2% in group II, 96.7% in group III, and 89.9% in group IV (P=0.093). The overall incidence of intraoperative complications was 11.8%. According to the modified Satava classification system, 6.1% of patients had grade 1, 5.1% had grade 2, and 0.54% had grade 3 complications. Intraoperative complications developed in 12.1% of patients in group I, 12.6% of patients in group II, 7.7% of patients in group III, and 12.2% of patients in group IV (P=0.625). Our findings clearly demonstrate that ureteroscopic treatment of ureteral stones can be safely and effectively performed with no use of contrast study imaging, except in doubtful cases of anatomical abnormalities.

Caracterização do mercado consumidor de "água aromatizada": hábitos e motivações para o consumo

O produto preparado líquido aromatizado, comercialmente conhecido como "água aromatizada" e recém-lançado no mercado brasileiro, é vendido com o apelo de saúde e inovação. No entanto, por se tratar de um produto pouco familiar aos hábitos de consumo da população brasileira, as motivações para o seu consumo ainda são desconhecidas. Diante deste contexto, o presente estudo teve por objetivo avaliar o perfil dos consumidores da cidade mineira de Juiz de Fora quanto ao produto, por meio de uma pesquisa de mercado realizada em junho de 2007. Para o levantamento dos dados, utilizou-se uma pesquisa descritiva direta e estruturada (survey) com 303 consumidores, por meio da aplicação de questionários estruturados. A faixa etária predominante (40,3%) situou-se entre 20 e 30 anos, sendo a maioria do sexo feminino (65%), com elevada escolaridade e renda familiar entre 1 e 6 salários mínimos (52,8%). Dos indivíduos entrevistados, 37% disseram que consumiram o produto motivados principalmente pelo fator novidade. Destes, 20,5% confundiram o produto com refrigerantes de baixa gaseificação. A maioria dos respondentes demonstrou desconhecer o produto, fato que pode estar relacionado a falta de divulgação nos estabelecimentos comerciais e pelas empresas através da mídia local.

Atividade antimicrobiana e antioxidante do óleo essencial de ho-sho (Cinnamomum camphora Ness e Eberm Var. Linaloolifera fujita)

Este trabalho teve como objetivo determinar a atividade antimicrobiana e antioxidante do óleo essencial de Ho-Sho. O principal componente do óleo essencial obtido a partir de folhas da planta submetidas ao processo de hidrodestilação foi o linalol (80 a 95% m/m). O óleo essencial mostrou atividade antimicrobiana para todos os microrganismos testados, com exceção de Pseudomonas aeruginosa. A maior atividade antimicrobiana do óleo essencial sobre as bactérias testadas foi observada sobre Xanthomonas campestris (33,0 mm) e a menor sobre Yersinia enterocolitica (10,5 mm). Para a concentração inibitória mínima (CIM), observou-se que todos os microrganismos apresentaram-se susceptíveis ao óleo essencial de Ho-Sho. A variação das CIM para as bactérias Gram-positivas foi de 1,00 mg.mL-1 (Streptococcus mutans) a 1,75 mg.mL-1 (Staphylococcus epidermidis). Já a variação das CIM para as bactérias Gram-negativas foi de 0,625 mg.mL-1 (Citrobacter freundii) a 2,50 mg.mL-1 (Shigella flexneri). Os resultados obtidos na determinação da atividade antioxidante do óleo essencial demonstram que o percentual antioxidante aumenta proporcionalmente à concentração de óleo essencial adicionado, atingindo o valor máximo de 97,49% de atividade antioxidante para a concentração de 50000 μg.mL-1.

Density and rheological parameters of goat milk

The rheological behavior and density of goat milk was studied as a function of solids concentration (10.5 to 50.0%) and temperature (273 to 331 k). Newtonian behavior was observed for values of total solids (TS) between 10.5 and 22.0% and temperatures from 276 to 331 k changing to pseudoplastic behavior without yield stress for TS from 25.0 to 39.4% at the same range of temperature. Goat milk with TS between 44.3 to 50.0% and temperatures of 273 to 296 k showed yield stress in addition to pseudoplastic behavior. At 303 to 331 k the power law model was observed again, without yield stress. The density of goat milk ranged from 991.7 to 1232.4 kg.m-3.

Nutritional quality and bioactive compounds of partially defatted baru almond flour

This study aimed to investigate the nutritional quality and bioactive potential of partially defatted baru (Dipteryx alataVog.) almond flour (BAF). The flour’s proximate and mineral compositions, total phenolic, tocopherols and carotenoids contents, antioxidant capacity, trypsin inhibitor and amino acid analyses were performed. An experiment was conducted with 24 male Wistar rats in order to evaluate the flour’s protein quality. BAF has high protein, fiber and mineral contents (iron, zinc, magnesium and copper), and it is a source of calcium. BAF presented relevant amounts of total phenolics (625 mg/100g) and good antioxidant capacity (130 µmol/Trolox eq). Autoclaved BAF showed essential amino acids profile, digestibility and protein quality better than in natura BAF. Autoclaved BAF might be used for human consumption as a source of quality protein and bioactive compounds, in healthy diets and processed foods.

Peritonites em pacientes em diálise peritoneal: análise de um único centro brasileiro segundo a Sociedade Internacional de Diálise Peritoneal

INTRODUÇÃO: A peritonite continua sendo a maior complicação para os pacientes em diálise peritoneal (DP). OBJETIVO: Este estudo teve como objetivo determinar as taxas de peritonite por episódio/ano (ep./ ano), ep./ano por microrganismo causador e pela mediana do número de peritonites nos pacientes em diálise peritoneal do Serviço de Nefrologia do Hospital São Lucas da PUCRS. MÉTODOS: Estudo retrospectivo e descritivo, no qual a amostra foi composta de pacientes que fizeram diálise peritoneal no Serviço de Nefrologia do HSL no período de 1984 a agosto de 2012; foram considerados somente os que possuíam dados completos. RESULTADOS: Dos 427 pacientes analisados, 53,2% eram do sexo feminino, com idade média de 48,0 ± 19,9 anos, 13% (56) de diabéticos e 71,5% (303) dos pacientes realizavam seu próprio tratamento. Ocorreram 503 episódios de peritonite e 255 pacientes tiveram pelo menos uma peritonite. Staphylococcus coagulase negativo foi o microrganismo mais prevalente. As causas de saída de tratamento foram óbito, transplante renal e peritonite, com 34,4, 25,8 e 19,2%, respectivamente. A taxa de peritonite foi de 0,63 ep./ano e ep./ ano por microrganismo foi de 0,18 ep./ ano para Staphylococcus coagulase negativo, e de 0,12 ep./ano para Staphilococcus aureus e Gram negativos. A mediana da unidade foi de 0,41. CONCLUSÃO: A taxa de peritonite ep./ano, e a mediana dos pacientes estudados encontram-se dentro do mínimo preconizado, mas abaixo das metas sugeridas, assim como a caracterização de ep./ano por microrganismo.