Transfer of penicillin resistance between Neisseriae in microcosm

Horizontal gene transfer between commensal and pathogenic Neisseriae is the mechanism proposed to explain how pathogenic species acquire altered portions of the penA gene, which encodes penicillin binding protein 2. These changes resulted in a moderately penicillin-resistant phenotype in the meningococci, whose frequency of isolation in Spain increased at the end of the 1980s. Little has been published about the possibility of this gene transfer in nature or about its simulation in the laboratory. We designed a simple microcosm, formed by solid and liquid media, that partially mimics the upper human respiratory tract. In this microcosm, penicillin-resistant commensal strains and the fully susceptible meningococcus were co-cultivated. The efficiency of gene transfer between the strains depended on the phase of bacterial growth and the conditions of culture. Resistance of penicillin was acquired in different steps irrespective of the source of the DNA. The presence of DNase in the medium had no effect on gene transfer, but it was near zero when nicked DNA was used. Cell-to-cell contact or membrane blebs could explain these results. The analysis of sequences of the transpeptidase domain of PBP2 from transformants, and from donor and recipient strains demonstrated that the emergence of moderately resistant transformants was due to genetic exchange between the co-cultivated strains. Finally, mechanisms other than penA modification could be invoked to explain decreased susceptibility

Assessing the Paradox Between Transmitted and Acquired HIV Type 1 Drug Resistance Mutations in the Swiss HIV Cohort Study From 1998 to 2012.

BACKGROUND: Transmitted human immunodeficiency virus type 1 (HIV) drug resistance (TDR) mutations are transmitted from nonresponding patients (defined as patients with no initial response to treatment and those with an initial response for whom treatment later failed) or from patients who are naive to treatment. Although the prevalence of drug resistance in patients who are not responding to treatment has declined in developed countries, the prevalence of TDR mutations has not. Mechanisms causing this paradox are poorly explored. METHODS: We included recently infected, treatment-naive patients with genotypic resistance tests performed ≤1 year after infection and before 2013. Potential risk factors for TDR mutations were analyzed using logistic regression. The association between the prevalence of TDR mutations and population viral load (PVL) among treated patients during 1997-2011 was estimated with Poisson regression for all TDR mutations and individually for the most frequent resistance mutations against each drug class (ie, M184V/L90M/K103N). RESULTS: We included 2421 recently infected, treatment-naive patients and 5399 patients with no response to treatment. The prevalence of TDR mutations fluctuated considerably over time. Two opposing developments could explain these fluctuations: generally continuous increases in the prevalence of TDR mutations (odds ratio, 1.13; P = .010), punctuated by sharp decreases in the prevalence when new drug classes were introduced. Overall, the prevalence of TDR mutations increased with decreasing PVL (rate ratio [RR], 0.91 per 1000 decrease in PVL; P = .033). Additionally, we observed that the transmitted high-fitness-cost mutation M184V was positively associated with the PVL of nonresponding patients carrying M184V (RR, 1.50 per 100 increase in PVL; P < .001). Such association was absent for K103N (RR, 1.00 per 100 increase in PVL; P = .99) and negative for L90M (RR, 0.75 per 100 increase in PVL; P = .022). CONCLUSIONS: Transmission of antiretroviral drug resistance is temporarily reduced by the introduction of new drug classes and driven by nonresponding and treatment-naive patients. These findings suggest a continuous need for new drugs, early detection/treatment of HIV-1 infection.

Snail blocks the cell cycle and confers resistance to cell death

The Snail zinc-finger transcription factors trigger epithelial-mesenchymal transitions (EMTs), endowing epithelial cells with migratory and invasive properties during both embryonic development and tumor progression. During EMT, Snail provokes the loss of epithelial markers, as well as changes in cell shape and the expression of mesenchymal markers. Here, we show that in addition to inducing dramatic phenotypic alterations, Snail attenuates the cell cycle and confers resistance to cell death induced by the withdrawal of survival factors and by pro-apoptotic signals. Hence, Snail favors changes in cell shape versus cell division, indicating that with respect to oncogenesis, although a deregulation/increase in proliferation is crucial for tumor formation and growth, this may not be so for tumor malignization. Finally, the resistance to cell death conferred by Snail provides a selective advantage to embryonic cells to migrate and colonize distant territories, and to malignant cells to separate from the primary tumor, invade, and form metastasis.

Participação da radiologia nas perícias necroscópicas de baleados realizadas no Instituto Médico-Legal do Rio de Janeiro

OBJETIVO: Este trabalho aborda as conseqüências de laudos necroscópicos incompletos de baleados, nos casos em que não foi possível o uso de recursos radiológicos para localizar os projéteis de arma de fogo. MATERIAIS E MÉTODOS: Foram analisados 8.185 laudos necroscópicos do Instituto Médico-Legal Afrânio Peixoto, Rio de Janeiro, RJ, referentes à demanda total de cadáveres no período de 1º de janeiro a 31 de dezembro de 2001, dos quais 3.122 casos corresponderam a necropsias de baleados. RESULTADOS: Desses casos, 309 corpos foram sepultados contendo ainda, no seu interior, projéteis de arma de fogo, podendo suscitar futuras indagações judiciais. No mesmo período foram solicitadas 23 exumações, 12 delas com a finalidade de recolher projéteis. Foram calculados os gastos relacionados à realização de necropsias de baleados - R$ 996,85 - e custos alusivos à realização de exumações com a finalidade de recolher projéteis de arma de fogo - R$ 1.155,40, visando a estabelecer o montante financeiro que poderia ser poupado pelos cofres públicos, a ser alocado para finalidades outras, se a perícia médico-legal de baleados, no exame inicial, obtivesse sucesso. CONCLUSÃO: Os resultados permitiram concluir que todos os atos necroscópicos de baleados devem seguir protocolos específicos, uma vez que perícias incompletas exigem exumação posterior, com gastos adicionais desnecessários.

Stepwise emergence of azole, echinocandin and amphotericin B multidrug resistance in vivo in Candida albicans orchestrated by multiple genetic alterations.

OBJECTIVES: The objective of this study was to characterize the underlying molecular mechanisms in consecutive clinical Candida albicans isolates from a single patient displaying stepwise-acquired multidrug resistance. METHODS: Nine clinical isolates (P-1 to P-9) were susceptibility tested by EUCAST EDef 7.2 and Etest. P-4, P-5, P-7, P-8 and P-9 were available for further studies. Relatedness was evaluated by MLST. Additional genes were analysed by sequencing (including FKS1, ERG11, ERG2 and TAC1) and gene expression by quantitative PCR (CDR1, CDR2 and ERG11). UV-spectrophotometry and GC-MS were used for sterol analyses. In vivo virulence was determined in the insect model Galleria mellonella and evaluated by log-rank Mantel-Cox tests. RESULTS: P-1 + P-2 were susceptible, P-3 + P-4 fluconazole resistant, P-5 pan-azole resistant, P-6 + P-7 pan-azole and echinocandin resistant and P-8 + P-9 MDR. MLST supported genetic relatedness among clinical isolates. P-4 harboured four changes in Erg11 (E266D, G307S, G450E and V488I), increased expression of ERG11 and CDR2 and a change in Tac1 (R688Q). P-5, P-7, P-8 and P-9 had an additional change in Erg11 (A61E), increased expression of CDR1, CDR2 and ERG11 (except for P-7) and a different amino acid change in Tac1 (R673L). Echinocandin-resistant isolates harboured the Fks1 S645P alteration. Polyene-resistant P-8 + P-9 lacked ergosterol and harboured a frameshift mutation in ERG2 (F105SfsX23). Virulence was attenuated (but equivalent) in the clinical isolates, but higher than in the azole- and echinocandin-resistant unrelated control strain. CONCLUSIONS: C. albicans demonstrates a diverse capacity to adapt to antifungal exposure. Potentially novel resistance-inducing mutations in TAC1, ERG11 and ERG2 require independent validation.

Stable eusociality via maternal manipulation when resistance is costless.

In many eusocial species, queens use pheromones to influence offspring to express worker phenotypes. Although evidence suggests that queen pheromones are honest signals of the queen's reproductive health, here I show that queen's honest signalling can result from ancestral maternal manipulation. I develop a mathematical model to study the coevolution of maternal manipulation, offspring resistance to manipulation and maternal resource allocation. I assume that (i) maternal manipulation causes offspring to be workers against offspring's interests; (ii) offspring can resist at no direct cost, as is thought to be the case with pheromonal manipulation; and (iii) the mother chooses how much resource to allocate to fertility and maternal care. In the coevolution of these traits, I find that maternal care decreases, thereby increasing the benefit that offspring obtain from help, which in the long run eliminates selection for resistance. Consequently, ancestral maternal manipulation yields stable eusociality despite costless resistance. Additionally, ancestral manipulation in the long run becomes honest signalling that induces offspring to help. These results indicate that both eusociality and its commonly associated queen honest signalling can be likely to originate from ancestral manipulation.

Net Neutrality: Measuring the Problem, Assessing the Legal Risks

Network neutrality is a growing policy controversy. Traffic management techniques affect not only high-speed, high-money content, but by extension all other content too. Internet regulators and users may tolerate much more discrimination in the interests of innovation. For instance, in the absence of regulatory oversight, ISPs could use Deep Packet Inspection (DPI) to block some content altogether, if they decide it is not to the benefit of ISPs, copyright holders, parents or the government. ISP blocking is currently widespread in controlling spam email, and in some countries in blocking sexually graphic illegal images. In 1999 this led to scrutiny of foreclosure of Instant Messaging and video and cable-telephony horizontal merger. Fourteen years later, there were in 2013 net neutrality laws implemented in Slovenia, the Netherlands, Chile and Finland, regulation in the United States and Canada , co-regulation in Norway, and self-regulation in Japan, the United Kingdom and many other European countries . Both Germany and France in mid-2013 debated new net neutrality legislation, and the European Commission announced on 11 September 2013 that it would aim to introduce legislation in early 2014. This paper analyses these legal developments, and in particular the difficulty in assessing reasonable traffic management and ‘specialized’ (i.e. unregulated) faster services in both EU and US law. It also assesses net neutrality law against the international legal norms for user privacy and freedom of expression

Calidad de vida y cáncer de próstata: un programa progresivo de fuerza-resistencia = Quality of life and prostate cancer: a progressive resistance strength program

This article presents the design and implementation of a progressive resistance strength program adapted to prostate cancer. The initial model corresponds to the guide of the American College Sports Medicine Position Stand (ACSM, 2009). This program includes the most habitual symptoms related to the illness and its treatments. The study design is quasi-experimental. The sample is 33 subjects in treatment phase. Study variables are tumour classification TNM, anthropometric measures, resistance strength, hypertension, fatigue, incontinence, pain and quality of life. After 24 weeks a significant improvement on resistance strength capacity is observed. This result is more consistent in lower extremities. Also improves hypertension, urinary incontinence, pain and quality of life. As conclusion, the improvement of the quality of life is mediated by the functional and physical capacity of the ill person

A utilização de imagens na identificação humana em odontologia legal

Este artigo de revisão sistemática tem por objetivo citar os métodos de identificação humana por meio da radiologia, utilizados em odontologia legal. Para isso, realizou-se revisão de literatura com 19 trabalhos selecionados dentre 45 encontrados, após aplicação de critérios de inclusão. Há diversas técnicas radiológicas que podem ser utilizadas para auxiliar na identificação humana, tanto individual como geral, incluindo a determinação do gênero, do grupo étnico e, principalmente, da idade. A análise de radiografias e tomografias ante-mortem e post-mortem tornou-se uma ferramenta fundamental nos processos de identificação em odontologia legal, principalmente com o refinamento das técnicas adquiridas com o avanço da própria radio-logia e com a incorporação da informática. Conclui-se que a partir do conhecimento adequado dos métodos disponíveis, o profissional em odontologia legal pode optar pelo método que melhor preencha as características necessárias para o sucesso da identificação, tendo cuidado na aplicação correta da técnica e na interpretação precisa das informações obtidas.

Metabolic syndrome and nonalcoholic fatty liver disease: Is insulin resistance the link?

Metabolic syndrome (MetS) is a disease composed of different risk factors such as obesity, type 2 diabetes or dyslipidemia. The prevalence of this syndrome is increasing worldwide in parallel with the rise in obesity. Nonalcoholic fatty liver disease (NAFLD) is now the most frequent chronic liver disease in western countries, affecting more than 30% of the general population. NAFLD encompasses a spectrum of liver manifestations ranging from simple steatosis to nonalcoholic steatohepatitis (NASH), fibrosis and cirrhosis, which may ultimately progress to hepatocellular carcinoma. There is accumulating evidence supporting an association between NAFLD and MetS. Indeed, NAFLD is recognized as the liver manifestation of MetS. Insulin resistance is increasingly recognized as a key factor linking MetS and NAFLD. Insulin resistance is associated with excessive fat accumulation in ectopic tissues, such as the liver, and increased circulating free fatty acids, which can further promote inflammation and endoplasmic reticulum stress. This in turn aggravates and maintains the insulin resistant state, constituting a vicious cycle. Importantly, evidence shows that most of the patients developing NAFLD present at least one of the MetS traits. This review will define MetS and NAFLD, provide an overview of the common pathophysiological mechanisms linking MetS and NAFLD, and give a perspective regarding treatment of these ever growing metabolic diseases.

Fatigue resistance of denture teeth

AIMS: There is no standard test to determine the fatigue resistance of denture teeth. With the increasing number of patients with implant-retained dentures the mechanical strength of the denture teeth requires more attention and valid laboratory test set-ups. The purpose of the present study was to determine the fatigue resistance of various denture teeth using a dynamic load testing machine. METHODS: Four denture teeth were used: Bonartic II (Candulor), Physiodens (Vita), SR Phonares II (Ivoclar Vivadent) and Trubyte (Dentsply). For dynamic load testing, first upper molars with a similar shape and cusp inclination were selected. The molar teeth were embedded in cylindrical steel molds with denture base material (ProBase, Ivoclar Vivadent). Dynamic fatigue loading was carried out on the mesio-buccal cusp at a 45° angle using dynamic testing machines and 2,000,000 cycles at 2Hz in water (37°C). Three specimens per group and load were submitted to decreasing load levels (at least 4) until all the three specimens no longer showed any failures. All the specimens were evaluated under a stereo microscope (20× magnification). The number of cycles reached before observing a failure, and its dependence on the load and on the material, has been modeled using a parametric survival regression model with a lognormal distribution. This allowed to estimate the fatigue resistance for a given material as the maximal load for which one would observe less than 1% failure after 2,000,000 cycles. RESULTS: The failure pattern was similar for all denture teeth, showing a large chipping of the loaded mesio-buccal cusp. In our regression model, there were statistically significant differences among the different materials, with SR Phonares II and Bonartic II showing a higher resistance than Physiodens and Trubyte, the fatigue resistance being estimated at around 110N for the former two, and at about 60N for the latter two materials. CONCLUSION: The fatigue resistance may be a useful parameter to assess and to compare the clinical risk of chipping and fracture of denture tooth materials.