Rho GTPase activating protein 21-mediated regulation of prostate cancer associated 3 gene in prostate cancer cell.

The overexpression of the prostate cancer antigen 3 (PCA3) gene is well-defined as a marker for prostate cancer (PCa) diagnosis. Although widely used in clinical research, PCA3 molecular mechanisms remain unknown. Herein we used phage display technology to identify putative molecules that bind to the promoter region of PCA3 gene and regulate its expression. The most frequent peptide PCA3p1 (80%) was similar to the Rho GTPase activating protein 21 (ARHGAP21) and its binding affinity was confirmed using Phage Bead ELISA. We showed that ARHGAP21 silencing in LNCaP prostate cancer cells decreased PCA3 and androgen receptor (AR) transcriptional levels and increased prune homolog 2 (PRUNE2) coding gene expression, indicating effective involvement of ARHGAP21 in androgen-dependent tumor pathway. Chromatin immunoprecipitation assay confirmed the interaction between PCA3 promoter region and ARHGAP21. This is the first study that described the role of ARHGAP21 in regulating the PCA3 gene under the androgenic pathway, standing out as a new mechanism of gene regulatory control during prostatic oncogenesis.

Genesis of People's Medicine Centre (PMC) for popularisation of generic medicine: A critical qualitative inquiry.

Introduction: The concerns of inaccessibility to essential medicines in India are well-studied and documented. Pradhan Mantri Bhartiya Janaushadhi Priyojana (PMBJP) is one of the policy initiatives to address the inaccessibility of essential medicine. Janaushadhi Kendra (People's Medicine Centre), which is part of PMBJP is being enquired in a limited way to understand its effectiveness. The province of Odisha has been chosen as the study area for the evaluation of People's Medicine Centres. Objective: The present study intends to inquire into the nature of People's Medicine Centre ownership, pharmacists' motivations and incentives to engage in business, perceived customers' trust and satisfaction, scheme beneficiaries, and challenges. Methods: A qualitative research approach has been adopted to evaluate the broader subjective accounts of the pharmacists and People's Medicine Centre. An open-ended interview guide was used. The topics of ownership, motivation, incentives, trust, satisfaction, perceived benefits, and challenges has been recorded from the participants. A total of seventeen in-depth interviews were conducted in the province of Odisha, India. Results: The study found that the ownership of People's Medicine Centre was of two types in the province of Odisha: public-NGO-owned People's Medicine Centres and public-private-owned People's Medicine Centres. The financial incentive provisions in the scheme attracted the private pharmacists. Pharmacists highlighted about the lower price of generic medicines compared to branded medicines, which is very popular among patients. They also pointed out that there is no difference in the efficacy of both medicines. The attitude of physicians, especially private physicians, were considered problematic for popularity and acceptance. Conclusion: The People's Medicine Centres in Odisha established themselves as trusted outlets despite physicians' unfavourable attitudes. Although the centres have not reached the required geographical coverage, economically developed regions have large number of centres, while backward regions have minimal presence. The scheme needs to be more conducive to the welfare of the masses living in remote and rural areas.

Peptide-based electrochemical biosensor for juvenile idiopathic arthritis detection.

Juvenile idiopathic arthritis (JIA) is a wide group of diseases, characterized by synovial inflammation and joint tissue damage. Due to the delay in the implementation of biomarkers into clinical practice and the association with severe sequels, there is an imperative need for new JIA diagnosis strategies. Electrochemical biosensors based on screen-printed electrodes and peptides are promising alternatives for molecular diagnosis. In this work, a novel biosensor for detecting juvenile idiopathic arthritis (JIA) was developed based on the immobilization of the PRF+1 mimetic peptide, as recognition biological element, on the surface of screen-printed carbon electrode. This biosensor was able to discriminate the JIA positive and negative serum samples from different individuals using differential pulse voltammetry, presenting limits of detection and quantification in diluted samples of 1:784 (v/v) and 1:235 (v/v), respectively. Evaluation by electrochemical impedance spectroscopy showed RCT 3 times higher for JIA positive sample than for a pool of human serum samples from healthy individuals. Surface analysis of the biosensor by atomic force microscopy, after contact with JIA positive serum, presented great globular clusters irregularly distributed. The long-term stability of the biosensor was evaluated, remaining functional for over 40 days of storage (after storage at 8°C). Therefore, a simple, miniaturized and selective biosensor was developed, being the first one based on mimetic peptide and screen-printed carbon electrode, aiming at the diagnosis of the juvenile idiopathic arthritis in real serum samples.

Automatic assignment of 1H-NMR spectra of small molecules.

A novel data-evaluation procedure for the automatic atom to peak or multiplet assignment of 1H-NMR spectra of small molecules has been developed using a fast and robust expert system. The applicability and reliability of the method are demonstrated by comparison of a manually assigned database of 1H-NMR spectra with the assignments produced by the automatic procedure. The results of this analysis show an excellent success ratio, indicating that this new algorithm can have a major impact as a time saving tool for the organic chemist. A new graphical feature used to illustrate both the stability and quality of the elementary assignments is also introduced.

Morfometria e análise populacional de colônias de Apoica flavissima (Hymenoptera, Vespidae) coletadas na Amazônia brasileira / Morphometric and population analysis of colonies of the Apoica flavissima (Hymenoptera, Vespidae) collected in Brazilian Amazonia

Two colonies of the social wasp Apoica flavissima were collected and analyzed to verify the population and morphometric averages of the individuals. Both colonies were in bushes at 700 mm above the floor, close to forest creeks. The colonies were at the beginning of their development stage and showed a reduced number of individuals, when compared with other studies on this genus. Means of adult parameters were: body length 21,6 mm ; width of thorax 3,8 mm; and width of head, 3,5 mm . Pupae presented these averages: body length 20,05 mm; width of thorax 3,75 mm ; and width of head 3,51 mm. Natural environment must offer good conditions for the development of wasps and their pattern nesting. Thus, high growth rates and different nesting behavior can demonstrate environmental influences on biology of this wasp group.

Duas colônias da vespa social Apoica flavissima foram coletadas e analisadas quanto aos parâmetros morfométricos e a população residente no ninho. Estas estavam fundadas a cerca de 700 mm do chão, nidificadas em plantas herbáceas, nas proximidades de igarapés. Ambas se encontravam no inicio do seu ciclo biológico e apresentavam um pequeno número de indivíduos, quando comparado com outros trabalhos com este gênero. Os adultos foram mensurados com médias de 21,62 mm de comprimento do corpo; 3,8 mm de largura máxima do tórax e 3,5 mm de largura da cabeça. As pupas apresentaram 20,05 mm de comprimento corpóreo; 3,75 mm de largura máxima do tórax e 3,51 mm de largura da cabeça. Ambientes naturais devem favorecer o crescimento dos indivíduos assim como manter os seus padrões de nidificação. Assim, uma discrepante taxa de crescimento e diferentes hábitos de nidificação, podem indicar a influência do ambiente sobre a biologia deste grupo.

Evaluation of hybrid treatments to produce high quality reuse water.

Four tertiary hybrid treatments to produce high quality reused water, fulfilling Brazilian drinking water regulations, from a slaughterhouse's secondary treated effluent were evaluated. The pilot plant with a capacity of 500 L h(-1) was set up and consisted of these stages: pre-filtration system (cartridge filter 50 micron, activated carbon filter, cartridge filter 10 micron), oxidation (H2O2) or second filtration (ceramic filter, UF) followed by UV radiation (90 L h(-1)). The best combination was T4: pre-filtration followed by H2O2 addition and UV radiation (AOP H2O2/UV). Disinfection kinetics by T4 followed pseudo first-order kinetics: k(T4) = 0.00943 s(-1) or 0.00101 cm2 mJ(-1). Three different zones (A, B, C) were observed in the UV254 degradation kinetics (pseudo-first order kinetics): k' decreased over time (k'(A) > k'(B) > k'(C)).

Prevalência e perfil de resistência a antimicrobianos de sorovares de Salmonella isolados de lingüiças suínas tipo frescal em Lages, SC / Prevalence and profile of resistance to antimicrobials of Salmonella serovars isolated from raw pork sausage in Lages, SC

The prevalence and profile of resistance to antimicrobials of Salmonella serovars isolated from raw pork sausage were studied in Lages county, Santa Catarina, Brazil. A total of 125 samples of 12 trademarks were collected in different commercial establishments. Salmonella sp. was present in 12.8 percent (16/125) of the samples and Typhimurium serovar was the most prevalent. Fourteen different antimicrobials were tested and most of the samples showed resistance to sulfonamide and tetracycline (81.2 percent). Eight positive samples (50 percent) were resistant at least to four antimicrobials, being considered as multi-resistant Salmonella. Seven (58.3 percent) trademarks were disagreement with the Brazilian law, representing a risk to the public health. The high level of resistance to the antimicrobials should produce a concern by the pig industry and veterinarians in order to prevent the transmission of resistant strains through the food chain.

Arteriocolic fistulae due to a stab wound: a rare cause of lethal lower gastrointestinal bleeding.

The authors describe the first reported case of lethal lower gastrointestinal bleeding due to an iliocolic fistulae following a single stab wound. They also discuss the pathophysiology of the enterovacscular fistulae, as well as a review of the literature and assess forensic characteristics. This unusual case underlines the importance of being aware of the victim's history in arteriocolic fistulae due to a stab wound.

Sudden death from a "broken heart" in a young woman.

A case of ruptured myocardial infarct in a young woman with no previous medical history is presented, as well as a review of the literature and forensic aspects are discussed.

Lethal paradoxical cerebral vein thrombosis due to suspicious anticoagulant rodenticide intoxication with chlorophacinone.

Superwarfarin exposure is a growing health problem, described in many countries. The authors report a case of suspicious chlorophacinone poisoning with a problematic diagnosis. They review the literature and discuss particularities of anticoagulant rodenticide intoxication, as well as the apparent contradiction between anticoagulant intoxication and lethal thrombosis.

Fatal and non-fatal bilateral delayed carotid artery dissection after manual strangulation.

Carotid artery (CA) dissection and/or thrombosis may occur in a number of medical and forensic relevant situations. However, post traumatic carotid artery dissection is considered rare, and often underestimated due to possible delayed presentation. We describe two cases of carotid artery dissection following strangulation with delayed symptoms as well as discuss medical diagnostic problems and their forensic implications.

Eficiência bionutricional de animais da raça Nelore, F1s Valdostana-Nelore e de mestiços de raças européias adaptadas / Bio-nutritional efficiency of Nellore, Valdostana-Nellore F1's and European adapted crossbred animals

Foram avaliados cinco grupos genéticos quanto à eficiência bionutricional (índice resultante da primeira função canônica de uma análise bivariada que envolveu ganho de peso e consumo de matéria seca) sendo seis animais da raça Nelore (N), cinco 1/2Romosinuano-1/2Caracu (RSC), quatro 1/2Senepol-1/2Caracu (SEC), quatro 1/2Belmont Red-1/2Caracu (BRC) e sete 1/2Valdostana-1/2Nelore (VAN). Não foi observada diferença significativa no desempenho dos animais mestiços Caracu, cuja média de eficiência bionutricional foi 519,8. O Nelore apresentou a menor eficiência bionutricional, 425,07, e os animais F1 Valdostana-Nelore, 499,30. Tendência semelhante foi observada para o ganho de peso. Os mestiços que envolveram raças européias adaptadas (RSC, SEC, BRC) apresentaram média de ganho de peso de 1,69kg/dia, enquanto os F1s Valdostana-Nelore alcançaram 1,57kg/dia e os da raça Nelore 1,29kg/dia. Não houve diferença no consumo diário de matéria seca entre os grupos genéticos, média de 7,76kg de MS ingerida/dia. Os grupos genéticos apresentaram conversão alimentar semelhante, com média de 4,87kg de MS ingerida/kg de ganho de peso.

[Inductively coupled plasma and clinical biology. Toxicological applications]. / Intérêt de la torche à plasma en biologie clinique. Quelles applications en toxicologie?

The multi-elementary quantitation method using inductively coupled plasma mass spectrometry has been widely developed for use with biological fluids. Many elements can be quantified simultaneously in biological fluids, including: Li, Be, B, Al, Mn, Co, Ni, Cu, Zn, Ga, Ge, As, Se, Rb, Sr, Mo, Pd, Cd, Sn, Sb, Te, Ba, W, Pt, Hg, Tl, Pb, Bi, U. The validation procedure is described by the French Society of Clinical Biology. Results for urine are corrected after creatinine determination. We report applications in clinical toxicology and forensic toxicology. Advances in inductively coupled plasma mass spectrometry in the field of clinical biology are particularly important for toxicological analysis. This powerful tool is helpful for better patient care and for the search for cause of death.

Fatal overdoses of tramadol: is benzodiazepine a risk factor of lethality?

Tramadol is a centrally acting analgesic agent used in the treatment of mild to moderate pain. It has a low affinity to opioid receptors and inhibits the reuptake of norepinephrin and serotonin producing an analgesic action by blocking nociceptive impulses in the spine. Although 21 drug-combined fatalities including tramadol have been reported, only two fatal overdoses in adults with tramadol alone have been reported to date. We report four additional lethal intoxications, assess the toxicity of tramadol, the detection method and the possible interaction with other central nervous system (CNS) depressants, particularly benzodiazepines. Similarities between tramadol and buprenorphine are discussed, and a possible cytochrome P450-based interaction between tramadol and benzodiazepine is considered. To our knowledge, this relationship has never been reported in the literature.

Bauhinia bauhinioides plasma kallikrein inhibitor: interaction with synthetic peptides and fluorogenic peptide substrates related to the reactive site sequence.

A serine proteinase inhibitor was purified from Bauhinia bauhinioides seeds after extraction with 0.15M NaCl by ion-exchange column chromatography on DEAE-Sephadex, gel filtration on Superose 12 column, Mono Q chromatography or alternatively by affinity chromatography on trypsin- Sepharose. The inhibitor is a single polypeptide chain with molecular mass 20 kDa by gel filtration on Superose 12, but was resolved into two peaks by ion - exchange chromatography on Mono Q (FPLC system). The main eluted peak inhibits trypsin (Ki = 0.6 nM), plasma kallikrein (Ki = 0.35 nM), plasmin (Ki = 33.1 nM), and weakly chymotrypsin (Ki = 2,700 nM), being the most effective plasma kallikrein inhibitor isolated from Bauhinia seeds. Therefore, it was denominated Bauhinia bauhinioides kallikrein inhibitor (BbKI). Activity is thermolabile and on trypsin inhibition optimum pH is 8.0. BbKI displays high homology to other plant Kunitz inhibitors, except for the absence of disulfide bridges, and the only cysteine residue is at the C-terminal position (residue 154) characterizes a distinct member of the Kunitz family. The affinity of the inhibitor to trypsin was confirmed by adsorption to trypsin-Sepharose resin and by isolation of the trypsin-inhibitor complex by gel filtration. Peptides with variations around the reactive site of BbKI (GLPVRFESPLRINIIKESY) were synthesized containing a quenched fluorogenic group. Trypsin but not plasma kallikrein substrates, these peptides strongly inhibited plasma kallikrein.

Synthetic peptides and fluorogenic substrates related to the reactive site sequence of Kunitz-type inhibitors isolated from Bauhinia: interaction with human plasma kallikrein.

We have previously described Kunitz-type serine proteinase inhibitors purified from Bauhinia seeds. Human plasma kallikrein shows different susceptibility to those inhibitors. In this communication, we describe the interaction of human plasma kallikrein with fluorogenic and non-fluorogenic peptides based on the Bauhinia inhibitors' reactive site. The hydrolysis of the substrate based on the B. variegata inhibitor reactive site sequence, Abz-VVISALPRSVFIQ-EDDnp (Km 1.42 microM, kcat 0.06 s(-1), and kcat/Km 4.23 x 10(4) M(-1) s(-1)), is more favorable than that of Abz-VMIAALPRTMFIQ-EDDnp, related to the B. ungulata sequence (Km 0.43 microM, kcat 0.00017 s(-1), and kcat/Km 3.9 x 10(2) M(-1) s(-1)). Human plasma kallikrein does not hydrolyze the substrates Abz-RPGLPVRFESPL-EDDnp and Abz-FESPLRINIIKE-EDDnp based on the B. bauhinioides inhibitor reactive site sequence, the most effective inhibitor of the enzyme. These peptides are competitive inhibitors with Ki values in the nM range. The synthetic peptide containing 19 amino acids based on the B. bauhinioides inhibitor reactive site (RPGLPVRFESPL) is poorly cleaved by kallikrein. The given substrates are highly specific for trypsin and chymotrypsin hydrolysis. Other serine proteinases such as factor Xa, factor XII, thrombin and plasmin do not hydrolyze B. bauhinioides inhibitor related substrates.