Genetic diversity and structure in two epiphytic orchids from the montane forests of southern Ecuador: The role of overcollection on Masdevallia rosea in comparison with the widespread Pleurothallis lilijae.

Ecuador has a high diversity of orchids, but little is known about levels of genetic diversity for the great majority of species. Understanding how orchids might adapt to changes is crucial as deforestation and fragmentation of forest ecosystems threaten the survival of many epiphytic orchids that depend on other species, such as fungi and their host trees, for germination, growth, and establishment. Overcollection and the illegal trade are also major concerns for the survival of wild populations of orchids. Despite increasing awareness, effective interventions are often limited by a lack of data concerning the impacts that overexploitation might have. To fill this gap, we tested the effects of overcollection in the genetic diversity and structure of Masdevallia rosea, a narrow distributed epiphytic orchid historically collected in Ecuador, in comparison with the widely distributed Pleurothallis lilijae. Genotyping based on AFLPs showed reduced levels of diversity in wild populations but most especially in the overcollected, M. rosea. Overall, genetic admixture was high in P. lilijae segregating populations by altitude levels while fewer genetic groups were found in M. rosea. Genetic differentiation was low in both species. A spatial genetic structure was found in P. lilijae depending on altitude levels, while no spatial genetic structure was found in M. rosea. These results suggest different scenarios for the two species: while gene flow over long distance is possible in P. lilijae, the same seems to be unlikely in M. rosea possibly due to the low levels of individuals in the known populations. In situ and ex situ conservation strategies should be applied to protect the genetic pool in these epiphytic orchid species, and to promote the connectivity between wild populations. Adopting measures to reduce overexploitation and to understand the impacts of harvesting in wild populations are necessary to strengthen the legal trade of orchids.

Transgene-induced cell death following dengue-2 virus infection in Aedes aegypti.

Dengue viruses (DENVs) are mosquito-borne flaviviruses causing millions of human infections each year and pose a challenge for public health systems worldwide. Aedes aegypti is the principal vector species transmitting DENVs to humans. Controlling Ae. aegypti is difficult due to the abundance of breeding sites and increasing insecticide resistance in the vector populations. Developing new vector control strategies is critical for decreasing the disease burden. One potential approach is genetically replacing Ae. aegypti populations with vector populations highly resistant to DENV transmission. Here, we focus on an alternative strategy for generating dengue 2 virus (DENV-2) resistance in genetically-modified Ae. aegypti in which the mosquitoes express an inactive form of Michelob_x (Mx), an antagonist of the Inhibitor of Apoptosis (IAP), to induce apoptosis in those cells in which actively replicating DENV-2 is present. The inactive form of Mx was flanked by the RRRRSAG cleavage motif, which was recognized by the NS2B/NS3 protease of the infecting DENV-2 thereby releasing and activating Mx which then induced apoptosis. Our transgenic strain exhibited a significantly higher mortality rate than the non-transgenic control when infected with DENV-2. We also transfected a DNA construct containing inactive Mx fused to eGFP into C6/36 mosquito cells and indirectly observed Mx activation on days 3 and 6 post-DENV-2 infections. There were clear signs that the viral NS2B/NS3 protease cleaved the transgene, thereby releasing Mx protein into the cytoplasm, as was confirmed by the detection of eGFP expression in infected cells. The present study represents proof of the concept that virus infection can be used to induce apoptosis in infected mosquito cells.

Pollination in the Rainforest: Scarce Visitors and Low Effective Pollinators Limit the Fruiting Success of Tropical Orchids.

A single plant might be visited by many flower visitors but not all might act as pollinators. Legitimate pollinators might also differ considerably in their efficiency, limiting pollination success. Unsuitable climatic conditions such as rain also affect pollinator activity. However, in the evergreen rainforest there is no prolonged dry season and flowering occurs usually under rain. Here, we explore the dependence on pollinators and the efficiency of flower visitors for the fruiting success of 10 Andean rainforest orchids. All species were self-compatible but strictly pollinator-dependent. Overall, we found low levels of fruit set in control flowers while experimental geitonogamous and cross-pollinations increased fruit set, revealing extensive pollination limitation in all populations. Seed viability dropped considerably after self and geitonogamous pollinations suggesting the possibility of early-acting inbreeding depression. Even though we monitored flower visitors on an extensive survey, few visitors were seen in these species and even fewer acted as legitimate pollinators. Thus, even though orchid pollination might be extremely diversified, these results show that few visitors are pollinating these species, explaining the low levels of fruit set recorded in the area studied.

Visceral obesity is not correlated with lymph node metastases nor Colorectal cancer survival / Obesidade visceral não está correlacionada a metastases nos linfonodos ou cancer coloretal

ABSTRACT Introduction: The presence of lymph node metastasis in colorectal cancer is determinant for prognosis and for treatment planning. The relationship between visceral fat and the prognosis is not fully documented in the literature, so this study intended to evaluate whether there is a relationship between the presence of visceral obesity and the presence of lymph node metastases and the prognosis of patients with colorectal cancer. Materials and methods: A sample of 68 patients who underwent surgery for colorectal cancer at Hospital de Braga between 1/1/2007 and 31/12/2007 was constructed, and their clinical and pathological data were recorded. Visceral fat, subcutaneous, and total fat areas were measured on preoperative computed tomography. Visceral obesity was defined as a ratio of visceral fat to total fat area >0.29. The ratio of metastatic lymph node (; number of metastatic lymph node/number of lymph node examined) was calculated. Results: There was a significant association between visceral obesity and male sex (p = 0.032). Patient survival at 5 and 10 years of follow-up was higher in patients with subcutaneous obesity in both periods, but not significant. There was a significant association between the ratio of metastatic lymph node and survival at 5 and 10 years (p = 0.03 and p = 0.002, respectively), with higher survival when ratio of metastatic lymph node = 0% and worse for ≥ 18%. Conclusion: In this study, no significant association was observed between visceral obesity and the number of metastatic lymph node, nor with survival at 5 and 10 years.

RESUMO Introdução: A presença de metastização ganglionar no câncer colorretal é determinante como fator de prognóstico e para planear o tratamento. A relação entre a presença de gordura visceral e o prognóstico não está totalmente documentada na literatura. Assim, pretende-se avaliar a existência de relação entre obesidade visceral e a presença de metástases ganglionares e o prognóstico de doentes com câncer colorretal. Materiais e métodos: Construiu-se uma amostra de 68 doentes operados por câncer colorretal no Hospital de Braga, entre 1/1/2007 e 31/12/2007, e registaram-se os seus dados clínico-patológicos e de seguimento. As áreas de gordura visceral, gordura subcutânea e gordura total foram medidas na tomografia computorizada pré-operatória. Obesidade visceral foi definida como um razão da gordura visceral relativamente à área total de gordura >0,29. Calculou-se a razão de linfonodos metastizados. Resultados: Verificou-se uma associação significativa entre obesidade visceral e sexo masculino (p = 0,032). A sobrevida dos pacientes, aos 5 e 10 anos de seguimento, foi superior naqueles com obesidade subcutânea em ambos períodos, contudo não significativa. Verificou-se uma associação significativa entre a sobrevivência em função da razão de linfonodos metastizados, aos 5 e 10 anos (p = 0,03 e p = 0,002; respectivamente), com maior sobrevivência quando a razão de linfonodos metastizados = 0% e pior quando ≥ 18%. Conclusão: Neste estudo não se observou uma associação significativa entre a obesidade visceral e o número de linfonodos metastizados nem com a sobrevida aos 5 e 10 anos.

Epidemiología molecular de aislados de Acinetobacter baumannii resistentes a carbapenems en Argentina / Molecular epidemiology of carbapenem-resistant isolates of Acinetobacter baumannii in Argentina

Se estudiaron 100 aislados consecutivos y no epidemiológicamente relacionados de Acinetobacter baumannii resistentes a los carbapenems, recuperados entre enero y agosto de 2016 de muestras clínicas en 11 hospitales de 10 provincias de la Argentina, ubicadas en distintas regiones del país. Los genes que codifican las carbapenemasas de Ambler clase D y clase B se investigaron mediante la técnica de PCR utilizando cebadores específicos. Todos los aislados se agruparon mediante las técnicas de 3-locus sequence typing y la secuenciación del gen blaOXA-51-like. El gen blaOXA-23 se recuperó en todos los aislados estudiados. La población de A. baumannii resistente a carbapenems en Argentina estuvo asociada, principalmente, con ST1 (45%), ST25 (34%) y ST79 (15%). ST25 se recuperó en todas las regiones estudiadas y no se detectó CC2.

One hundred sequential, epidemiologically unrelated carbapenem-resistant- Acinetobacter baumannii isolates from 11 hospitals in 10 Argentine provinces were collected between January and August 2016. Genes coding for Ambler class D and B carbapenemases were investigated by PCR using specific primers. All isolates were typed using the 3-locus sequence typing and b/aOXA-51-like sequence-based typing techniques. The blaOXA-23 gene was recovered in all isolates studied. The population of carbapenem-resistant- A. baumannii in Argentina was principally associated with ST1 (45%), ST25 (34%) and ST79 (15%). ST25 was recovered in all the regions studied and CC2 was not detected.

Características clínicas y epidemiológicas de pacientes con paracoccidioidomicosis diagnosticados en un hospital de Resistencia, Chaco / Clinical and epidemiological characteristics of patients with paracoccidioidomycosis diagnosed in a hospital of Resistencia, Chaco

Con el objetivo de describir las características clínico-epidemiológicas de la paracoccidioidomicosis, se realizó un estudio descriptivo de los casos diagnosticados por el Servicio de Microbiología Clínica del hospital de adultos Dr. Julio C. Perrando, de la ciudad de Resistencia (Chaco, Argentina). Entre 2011 y 2014 se detectaron 46 casos. En 2013 y 2014 se constató un incremento de la tasa de incidencia de alrededor de 4 veces con respecto a los anos anteriores. La forma crónica fue la predominante, con una media de edad de los pacientes de 53 anos. Del total de ellos, a 39 se les realizaron pruebas serológicas. En 15 de 39 casos, las pruebas serológicas fueron la única herramienta diagnóstica, mientras que en 4 de estos casos con diagnóstico microbiológico, la prueba resultó no reactiva. La inclusión de la paracoccidioidomicosis en el diagnóstico diferencial de pacientes de áreas endémicas que presentan un síndrome infeccioso inespecífico y la aplicación de las herramientas diagnósticas disponibles contribuyen al diagnóstico oportuno, así como a disminuir las secuelas de esta afección y su impacto socioeconómico.

In order to describe the clinical and epidemiological characteristics of paracoccidioidomycosis, a descriptive study of all the cases diagnosed by the Clinical Microbiology Service at Dr. Julio C. Perrando hospital in the city of Resistencia (Chaco Province, Argentina) was conducted. Between 2011 and 2014, 46 cases were detected. In the period 2013-2014, an almost 4-fold increase in the incidence rate was detected. The chronic form of the disease was predominant with an average age of 53 years. Serological tests in 39 out of 46 patients were performed. In 15 of 39 patients, serological tests were the only diagnostic tool while in 4 patients with a microbiological diagnosis serological tests were non-reactive. In patients from endemic areas with non-specific infectious syndrome it is important to include paracoccidioidomycosis in the differential diagnosis and to apply all available diagnostic tools to reach a timely diagnosis and to reduce the long-term sequelae and their socio-economic impact.

Rectal carcinoid tumor: diagnosis and management / Tumor carcinoide de reto: diagnóstico e orientação

ABSTRACT The authors present a case of rectal carcinoid tumor in an asymptomatic patient who sought a coloproctology service with the purpose of colorectal cancer prevention. During colonoscopy, a polyp lesion was found in the rectum, and it was resected with a polypectomy loop. Anatomopathological examination revealed a rectal carcinoid tumor with compromised margins. The lesion site was resected again and pathological exam no longer showed neoplasia. A systematic review of the issue was performed, discussing diagnosis aspects, difficulties in the choice of therapeutic approaches, and prognosis. The conclusion is that the rarity of the disease brings difficulties in the choice of treatment; although it has a good prognosis in most cases, its malignant potential cannot be underestimated.

RESUMO Apresentamos um caso clínico de um tumor carcinóide de reto em um paciente assintomático que procurou um serviço de colo-proctologia com o objetivo de fazer prevenção ao câncer colorretal. Durante a colonoscopia observou-se uma lesão polipóide no reto, a qual foi ressecada com alça de polipectomia. O exame anatomopatológico evidenciou tumor carcinóide de reto com comprometimento de margem. O local foi novamente ressecado e o resultado patológico não mais evidenciou neoplasia. Neste artigo é feita uma revisão bibliográfica do assunto abordando aspectos diagnósticos, dificuldade na escolha da conduta terapêutica e prognóstica. O trabalho conclui que a raridade do caso implica em dificuldades na escolha da conduta e que apesar de ter bom prognóstico, em uma parcela dos casos o seu potencial maligno não deve ser desprezado.

[Clinical and epidemiological characteristics of patients with paracoccidioidomycosis diagnosed in a hospital of Resistencia, Chaco]. / Características clínicas y epidemiológicas de pacientes con paracoccidioidomicosis diagnosticados en un hospital de Resistencia, Chaco.

In order to describe the clinical and epidemiological characteristics of paracoccidioidomycosis, a descriptive study of all the cases diagnosed by the Clinical Microbiology Service at Dr. Julio C. Perrando hospital in the city of Resistencia (Chaco Province, Argentina) was conducted. Between 2011 and 2014, 46 cases were detected. In the period 2013-2014, an almost 4-fold increase in the incidence rate was detected. The chronic form of the disease was predominant with an average age of 53 years. Serological tests in 39 out of 46 patients were performed. In 15 of 39 patients, serological tests were the only diagnostic tool while in 4 patients with a microbiological diagnosis serological tests were non-reactive. In patients from endemic areas with non-specific infectious syndrome it is important to include paracoccidioidomycosis in the differential diagnosis and to apply all available diagnostic tools to reach a timely diagnosis and to reduce the long-term sequelae and their socio-economic impact.

[Molecular epidemiology of carbapenem-resistant isolates of Acinetobacter baumannii in Argentina]. / Epidemiología molecular de aislados de Acinetobacter baumannii resistentes a carbapenems en Argentina.

One hundred sequential, epidemiologically unrelated carbapenem-resistant- Acinetobacter baumannii isolates from 11 hospitals in 10 Argentine provinces were collected between January and August 2016. Genes coding for Ambler class D and B carbapenemases were investigated by PCR using specific primers. All isolates were typed using the 3-locus sequence typing and blaOXA-51-like sequence-based typing techniques. The blaOXA-23 gene was recovered in all isolates studied. The population of carbapenem-resistant- A. baumannii in Argentina was principally associated with ST1 (45%), ST25 (34%) and ST79 (15%). ST25 was recovered in all the regions studied and CC2 was not detected.

Oviplate: A Convenient and Space-Saving Method to Perform Individual Oviposition Assays in Aedes aegypti.

Aedes aegypti is the principal vector of the urban arboviruses and the blood ingestion is important to produce the eggs in this species. To analyze the egg production in Ae. aegypti, researchers frequently use small cages or Drosophila vials to collect eggs from gravid females. Although it is affordable, the setup is time- and space-consuming, mainly when many mosquitoes need to be individually analyzed. This study presents an easy, cheap, and space-saving method to perform individual oviposition assays in Ae. aegypti using cell culture plates. This new method to access fecundity rate was named "oviplate". The oviplates are setup with 12- or 24-well plates, distilled water and filter paper and they are 78 to 88% cheaper than the traditional Drosophila vial assay, respectively. Furthermore, to allocate 72 vitellogenic females in an insectary using Drosophila vial is necessary 4100 cm³ against 1400 cm³ and 700 cm³ when using 12- and 24-well plates, respectively. No statistical differences were found between the number of eggs laid in Drosophila vials and the oviplates, validating the method. The oviplate method is an affordable, and time- and space-efficient device, and it is simpler to perform individual fecundity analyses in Ae. aegypti.

Mosquito pornoscopy: Observation and interruption of Aedes aegypti copulation to determine female polyandric event and mixed progeny.

Ades aegypti is the most important arbovirus vector in the world, and new strategies are under evaluation. Biological studies mentioning the occurrence of a second mate in Aedes aegypti can interfere with vector control program planning, which involves male mosquito release technique. This study presents different experiments to show the occurrence of mixed progeny. Mixed male crosses (using a combination of different type of males in confinement with virgin females) showed no polyandric female. Individual crosses with male substitution in every gonotrophic cycle also did not show any polyandric female. Individual crosses with a 20 minutes interval, with subsequent male change, showed that only a few females presented mixed offspring. The copulation breach in three different moments, group A with full coitus length, group B the coitus was interrupted in 5-7 seconds after the start; and group C, which the copulation was interrupted 3 seconds after started. In summary, group A showed a majority of unique progeny from the first male; group B showed the higher frequency of mixed offspring and group C with the majority of the crosses belonging to the second male. To conclude, the occurrence of a viable second mate and mixed offspring is only possible when the copulation is interrupted; otherwise, the first mate is responsible for mixed progeny.

Satisfação de médicos no trabalho da atenção primária à saúde / Satisfaction of physicians working in primary health care

Neste estudo avaliou-se a satisfação no trabalho dos médicos da Atenção Primária de uma capital do sul do Brasil. Foram entrevistados quinze médicos de Unidades de Saúde. As principais motivações citadas para atuarem na Atenção Primária foram a estabilidade do emprego público, a dificuldade de se estabelecer na iniciativa privada e a defesa do direito universal à saúde. As principais questões geradoras de insatisfação relacionaram-se ao elevado número de consultas, falta de respaldo dos níveis secundário e terciário de atenção à saúde, limitações da autonomia decisória, desgaste físico e emocional, baixa resolubilidade dos serviços, desvalorização perante aos colegas e opinião pública.

This study assessed work satisfaction among Primary Health Care doctors from a capital city in Southern Brazil. Fifteen physicians from Basic Health Units were interviewed. The main motivations cited for working in Primary Health Care were the stability of the public service job, the difficulty of establishing a career in the private sector, and the defense of the universal right to health access. The main points that generated dissatisfaction were related to the high number of appointments per working hour, lack of support from the secondary and tertiary health care levels, limited decision-making autonomy, elevated physical and emotional distress, low resolution capability of health services, and lack of esteem from colleagues and society.

In Vivo Paracoccidioides sp. Biofilm on Vascular Prosthesis.

Paracoccidioidomycosis is an endemic mycosis caused by Paracoccidioides species limited to Latin America arising with the chronic form in 90% of cases. The capacity of microorganisms to form biofilms is considered of great importance medical since can contribute to the persistence and to the chronic state of the diseases. The ability of Paracoccidioides to form biofilm has been demonstrated in vitro. In our study, for the first time we have observed this capability in vivo on a vascular prosthesis using scanning electron microscope showing a dense network of Paracoccidioides yeasts covered by an extracellular matrix.

Outcomes of extensive hybridization and introgression in Epidendrum (Orchidaceae): can we rely on species boundaries?

Hybridization has the potential to contribute to phenotypic and genetic variation and can be a major evolutionary mechanism. However, when hybridization is extensive it can also lead to the blurring of species boundaries and the emergence of cryptic species (i.e., two or more species not distinguishable morphologically). In this study, we address this hypothesis in Epidendrum, the largest Neotropical genus of orchids where hybridization is apparently so common that it may explain the high levels of morphological diversity found. Nonetheless, this hypothesis is mostly based on the intermediacy of morphological characters and intermediacy by itself is not a proof of hybridization. Therefore, in this study, we first assessed the existence of hybrids using cpDNA and AFLP data gathered from a large-scale sampling comprising 1038 plants of three species of Epidendrum (E. calanthum, E. cochlidium and E. schistochilum). Subsequently, a Bayesian assignment of individuals into different genetic classes (pure species, F1, F2 or backcross generations) revealed that hybrid genotypes were prevalent in all sympatric populations. In most cases, parental species were not assigned as pure individuals, rather consisting in backcrossed genotypes or F1 hybrids. We also found that reproductive barriers are apparently very weak in Epidendrum because the three species largely overlapped in their flowering periods and interspecific crosses always produced viable seeds. Further, hybridization contributed to enhance floral variability, genome size and reproductive success since we found that these traits were always higher in hybrid classes (F1, F2 and backcrosses) than in pure parental species, and offer an explanation for the blurring of species boundaries in this genus of orchids. We hypothesize that these natural hybrids possess an evolutionary advantage, which may explain the high rates of cryptic species observed in this genus.

SNP typing reveals similarity in Mycobacterium tuberculosis genetic diversity between Portugal and Northeast Brazil.

Human tuberculosis is an infectious disease caused by bacteria from the Mycobacterium tuberculosis complex (MTBC). Although spoligotyping and MIRU-VNTR are standard methodologies in MTBC genetic epidemiology, recent studies suggest that Single Nucleotide Polymorphisms (SNP) are advantageous in phylogenetics and strain group/lineages identification. In this work we use a set of 79 SNPs to characterize 1987 MTBC isolates from Portugal and 141 from Northeast Brazil. All Brazilian samples were further characterized using spolygotyping. Phylogenetic analysis against a reference set revealed that about 95% of the isolates in both populations are singly attributed to bacterial lineage 4. Within this lineage, the most frequent strain groups in both Portugal and Brazil are LAM, followed by Haarlem and X. Contrary to these groups, strain group T showed a very different prevalence between Portugal (10%) and Brazil (1.5%). Spoligotype identification shows about 10% of mis-matches compared to the use of SNPs and a little more than 1% of strains unidentifiability. The mis-matches are observed in the most represented groups of our sample set (i.e., LAM and Haarlem) in almost the same proportion. Besides being more accurate in identifying strain groups/lineages, SNP-typing can also provide phylogenetic relationships between strain groups/lineages and, thus, indicate cases showing phylogenetic incongruence. Overall, the use of SNP-typing revealed striking similarities between MTBC populations from Portugal and Brazil.

Hydrotherapy study in patients with failed back surgery syndrome: a therapeutic option / Estudo da hidroterapia em pacientes com síndrome pós-laminectomia: uma opção terapêutica

The study was carried out to evaluate the efficacy of hydrotherapy on ?failed back surgery syndrome?. Eight participants were assigned to an aquatic exercise group. The analgesic medication was withdrawn during the study. Evaluation parameters included the results of the Visual Analogue Scale (VAS) score and spine mobility. The patients were assessed at baseline and at the end of the 6-week follow-up period. The results showed that hydrotherapy had a positive impact on the outcome of chronic post-surgical low back pain. After 6 weeks of hydrotherapy, without analgesic medication, the overall mobility of the spine improved and back pain decreased. We conclude that aquatic exercises may positively impact the outcome of ?failed back surgery syndrome?, probably remaining one of the few successful measures in the management of this kind of patient.

O presente estudo foi realizado para avaliar a eficácia da hidroterapia na ?síndrome pós-laminectomia?. Oito participantes foram designados para um grupo de exercícios aquáticos. A medicação analgésica foi descontinuada durante o estudo. Os parâmetros de avaliação incluíram o escore da Escala Visual Analógica (EVA) e a mobilidade da coluna vertebral. Os pacientes foram avaliados em condições basais e ao final do período de seguimento de seis semanas. Os resultados mostraram que a hidroterapia teve um impacto positivo nos resultados da dor lombar crônica pós-cirúrgica. Após seis semanas de hidroterapia, sem medicação analgésica, a mobilidade geral da coluna melhorou e a dor lombar diminuiu. Concluímos que exercícios na água podem ter um impacto positivo no resultado da síndrome da falência da cirurgia lombar, provavelmente sendo uma das poucas medidas bem sucedidas no tratamento desse tipo de paciente.