Cryptic species in Pagamea coriacea sensu lato (Rubiaceae): evidence from morphology, ecology and reproductive behavior in a sympatric context

In this study we explore morphological and ecological variation in sympatric populations of Pagamea coriacea s.l. - a species complex from white-sand vegetation in the Amazon. A total of 147 trees were sampled and monitored at three nearby sites in Central Amazon, Brazil. Multivariate analyses of morphology indicated two distinct groups (A and B), which also differed in bark type, each containing subgroups associated with sexual dimorphism. However, a single hermaphroditic individual was observed within group B. As expected, all pistillate plants produced fruits, but 23% of the staminate plants of group B, and 5% of group A also produced fruits. This variation suggests that the sexual systems of both groups are between dioecy and gynodioecy. There was an overlap in flowering phases between the two groups, but the pattern of floral maturation differed. Ecologically, plants of group B were found in more shaded habitats and over sandstone bedrocks, while group A was prevalent in deeper sandy soils as canopy plants. The significances of morphological and environmental differences were tested by a multivariate analysis of variance, and a canonical discriminant analysis assessed the importance of variables. The coexistence in sympatry of two discrete morphological groups in the P. coriacea s.l., with different habitat preferences and reproductive behaviors, indicates they represent distinct species.

Reproductive aspects of Macrobrachium amazonicum (Decapoda: Palaemonidae) in the State of Amapá, Amazon River mouth

Macrobrachium amazonicum is an indigenous prawn vastly distributed in basins of South America, widely exploited by artisanal fisheries in northern and northeastern Brazil and, with great potential for aquaculture. This study aimed to investigate general aspects of population structure and reproductive characteristics (size at first maturity, fecundity and reproductive output) of M. amazonicum from two important areas to artisanal prawn fishing located at the mouth of the Amazon River, State of Amapá. The specimens were captured using 20 handcrafted traps called "matapi". A number of 5,179 prawns were captured, 2,975 females and 2,195 males resulting in 1.35:1 female to male ratio. Santana Island and Mazagão Velho showed females predominated in the population. A reproductive peak period was observed from January to April/2009 and in December/2010, coinciding with the period of higher rainfall. The recruitment peak occurred in June and July/2009. Egg-bearing females ranged in size (carapace length) from 11.10 to 29.6 mm. Fecundity increased with female size and reached up to 7,417 eggs. This amount of eggs is considered low if compared with other Macrobrachium estuarine species. Mean egg volume increased gradually from 0.121 to 0.24 mm³ during embryogenesis, representing 68.5% of overall increase from Stage I to Stage III. Eggs of M. amazonicum are small; this is typical for Macrobrachium species, which depends on brackish water to complete the larval development. Irrespective of female size, reproductive output of M. amazonicum varied between 4.8 and 21.85% of their body weight into eggs production.

Reproductive biology of Amasonia obovata Gleason (Laminaceae)

Floral mechanisms that ensure seed production via autogamy are more likely to occur in species growing in environments where pollination is scarce. Amasonia obovata was studied in the State of Mato Grosso-Brazil, from 2009 to 2012, to analyze the morphological and reproductive characteristics, aside from investigating the association of the reproductive success with the pollinator frequency and identity. The flowering and fruiting of A. obovata was concentrated in a period of five months during the rainy season. The dichogamy in flowers of A. obovata is not clearly defined, since the sexual functions were overlapped in the male and female phases. The species is self-compatible and not apomictic. The fruiting percentage obtained by hand self-pollination did not differ from cross-breeding (F = 0.74, P =0.39). In the observations from 2010 to 2012, a hummingbird (Thalurania furcata) legitimate visited 20-100% of the flowers in the male and female phases on different A. obovata plants. Due to the high frequency, this hummingbird was considered the single potential pollinator of the species. These findings show that a limited availability of pollinators may select for floral traits and plant mating strategies that lead to a system of self-fertilization.

Reproductive biology of Ameerega trivittata (Anura: Dendrobatidae) in an area of terra firme forest in eastern Amazonia

The reproductive success of tropical amphibians is influenced by factors such as body size and the characteristics of breeding sites. Data on reproductive biology are important for the understanding of population dynamics and the maintenance of species. The objectives of the present study were to examine the abundance of Ameerega trivittata, analyze the use of microhabitats by calling males and the snout-vent length (SVL) of breeding males and females, the number of tadpoles carried by the males and mature oocytes in the females, as well as the relationship between the SVL of the female and both the number and mean size of the mature oocytes found in the ovaries. Three field trips were conducted between January and September, 2009. A total of 31 plots, with a mean area of 2.3 ha, were surveyed, resulting in records of 235 individuals, with a mean density of 3.26 individuals per hectare. Overall, 66.1% of the individuals sighted were located in the leaf litter, while 17.4% were perched on decaying tree trunks on the forest floor, 15.7% on the aerial roots of Cecropia trees, and 0.8% on lianas. Males were observed transporting a mean of 10.8 tadpoles on their backs. A significant correlation was found between the size of the females and the mean diameter of the oocytes. New data were collected on the size of oocytes and no pattern was found in the type of perches used by calling males of the different Ameerega species.

Ex situ collection of Limonium spp.: cytogenetic and reproductive characterization for the conservation of rare species

Dissertação mestrado em Biologia Molecular, Biotecnologia e Bioempreendedorismo em Plantas

Reproductive biology of Macrobrachium surinamicum (Decapoda: Palaemonidae) in the Amazon River mouth

Macrobrachium surinamicum is an indigenous prawn distributed from the lower Amazon and Tocantins river basins to Venezuela in the Orinoco Delta region. It is common bycatch fauna of Macrobrachium amazonicum artisan fishing in the states of Pará and Amapá. The aim of this study was to investigate aspects on reproductive biology (reproductive period, size of sexual maturity population, fecundity, reproductive output and recruitment) of M. surinamicum from four important areas to artisanal prawn fishing located at the Amazon River mouth (Amapá and Pará). The specimens were captured using 20 handcrafted traps called "matapi". A number of 675 prawns were captured, 258 males, 409 females and eight juveniles, resulting in 1:1.6 (Male: Female) sex ratio. The reproductive peak period occurred from March to July, coinciding with the higher rainfall period. The juvenile prawn occurred only in May and July. Total length of egg-bearing females ranged from 12.12 to 38.30 mm, with mean female length at first maturity (L50) of 23.7 mm. Fecundity increased with prawn size and varied between 174 and 1780 eggs per female. Mean egg volume increased gradually from 0.031 (Stage I) to 0.060 mm³ (Stage III) during embryogenesis. Macrobrachium surinamicum depends on brackish water to complete the larval development. Irrespective of female size, reproductive output of M. surinamicum varied between 4.3 % and 35.5 % of their body weight for egg production. The knowledge of the reproductive biology reported in the present study is an important tool to define strategies to preserve M. surinamicum in Amazon River mouth.

“PhenoWorld”: a new multidimensional strategy for studying behaviour in rodents

Tese de Doutoramento em Ciências da Saúde

Percutaneous handling of coronary lesions >20mm through stents. Is there a first choice strategy?

OBJECTIVE - This study compared the early and late results of the use of one single stent with those of the use of multiple stents in patients with lesions longer than 20mm. METHODS - Prospective assessment of patients electively treated with stents, with optimal stent deployment and followed-up for more than 3 months. From February '94 to January '98, 215 patients with lesions >20mm were treated. These patients were divided into 2 groups as follows: Group A - 105 patients (49%) with one stent implanted; Group B - 110 patients (51%) with multiple stents implanted. RESULTS - The mean length of the lesions was 26mm in group A (21-48mm) versus 29mm in group B (21-52mm) (p=0.01). Major complications occurred in one patient (0.9%) in group A (subacute thrombosis, myocardial infarctionand death) and in 2 patients (1.8%) in group B (one emergency surgery and one myocardial infarction) (p=NS). The results of the late follow-up period (>6 months) were similar for both groups (group A = 82% vs group B = 76%; p=NS), and we observed an event-free survical in 89% of the patients in group A and in 91% of the patients in group B (p=NS). Angina (group A = 11% vs group B = 7%) and lesion revascularization (group A = 5% vs group B = 6%; p=NS) also occurred in a similar percentage. No infarction or death was observed in the late follow-up period; restenosis was identified in 33% and 29% of the patients in groups A and B, respectively (p=NS). CONCLUSION - The results obtained using one stent and using multiple stents were similar; the greater cost-effectiveness of one stent implantation, however, seems to make this strategy the first choice.

Efficacy of a diagnostic strategy for patients with chest pain and no ST-segment elevation in the emergency room

PURPOSE: To evaluate the efficacy of a systematic model of care for patients with chest pain and no ST segment elevation in the emergency room. METHODS: From 1003 patients submitted to an algorithm diagnostic investigation by probability of acute ischemic syndrome. We analyzed 600 ones with no elevation of ST segment, then enrolled to diagnostic routes of median (route 2) and low probability (route 3) to ischemic syndrome. RESULTS: In route 2 we found 17% acute myocardial infarction and 43% unstable angina, whereas in route 3 the rates were 2% and 7%, respectively. Patients with normal/non--specific ECG had 6% probability of AMI whereas in those with negative first CKMB it was 7%; the association of the 2 data only reduced it to 4%. In patients in route 2 the diagnosis of AMI could only be ruled out with serial CKMB measurement up to 9 hours, while in route 3 it could be done in up to 3 hours. Thus, sensitivity and negative predictive value of admission CKMB for AMI were 52% and 93%, respectively. About one-half of patients with unstable angina did not disclose objective ischemic changes on admission. CONCLUSION: The use of a systematic model of care in patients with chest pain offers the opportunity of hindering inappropriate release of patients with ACI and reduces unnecessary admissions. However some patients even with normal ECG should not be released based on a negative first CKMB. Serial measurement of CKMB up to 9 hours is necessary in patients with medium probability of AMI.

Mortality due to cardiovascular disease in women during the reproductive age (15 to 49 years), in the State of São Paulo, Brazil, from 1991 to 1995

OBJECTIVE: To describe mortality due to cardiovascular diseases in women during the reproductive age (15 to 49 years) in the state of São Paulo, Brazil, from 1991 to 1995. METHODS: A list of all deaths and their underlying causes, coded according to the International Classification of Diseases, 9th revision, multiple causes of death, and estimates of the female population according to age groups were provided by the SEADE Foundation. Specific coefficients for 100 thousand women for each year as well as the medians of these coefficients related to 5 years, and the percentage of death by subgroups were calculated. RESULTS: Cerebrovascular diseases have the highest coefficients (14.24 for 100 thousand females), followed by ischemic heart disease (7.37), other heart diseases (6.39), hypertensive disease (3.03), chronic rheumatic heart disease (1.58), pulmonary vascular diseases (1.29), and active rheumatic fever (0.05). Systemic arterial hypertension, as an associated cause, occurred in 55.3% to 57.8% of all the deaths due to intracerebral hemorrhage and in 30.4% to 30.8% due to subarachnoid hemorrhage. CONCLUSION: The significance of cerebrovascular diseases, coronary artery disease, and systemic arterial hypertension as causes of mortality suggests the need to emphasize preventive actions for young women who have the potential to reproduce to avoid possible complications in future pregnancies, and premature mortality.

Sistemática filogenética y biología reproductiva con énfasis en solanum y capsicum (solanaceae). Systematics, phylogeny and reproductive biology with emphasis in solanum and capsicum (solanaceae).

Muchas respuestas a preguntas básicas sobre relaciones evolutivas, ubicación sistemática y evolución de caracteres morfológicos y ecológicos pueden ser obtenidas a través de las reconstrucciones filogenéticas. Sobre este contexto se pretende encarar en este proyecto estudios de filogenia molecular, revisiones sistemáticas, biología reproductiva y citogenética en Solanáceas americanas. Se intentará resolver la delimitación específica de Solanum sect. Solanum y Geminata, y Capsicum, y establecer relaciones filogenéticas en estos grupos. Se harán revisiones analizándose caracteres vegetativos y reproductivos críticos para evaluar su variabilidad y definir su valor taxonómico; para los estudios moleculares se utilizarán los marcadores ndhF, trnT-L, trnL-F y waxy. En base a los resultados se propondrán agrupamientos y relaciones de parentesco. Además, se hará un estudio cariosistemático para caracterizar y circunscribir especies en Solanum y miembros de la tribu Physaleae, y hasta variedades y/o cultivares en Capsicum, mediante técnicas clásicas y de bandeos de fluorescencia y AgNOR e hibridación in situ fluorescente (FISH). A nivel reproductivo, se estudiará la ecofisiología en las estructuras masculinas y su incidencia en la fructificación en Capscium baccatum. El desarrollo de esta temática comprende experiencias in vivo (a campo y en laboratorio) así como estudios histológicos y químicos.Se espera avanzar en la resolución de algunos problemas: 1) la complicada delimitación de especies de los taxones en estudio; 2) las relaciones filogenéticas en algunos de ellos; 3) la falta de conocimiento de la organización genómica; 4) el origen de las especies cultivadas de Capsicum. En cuanto a la biología reproductiva, para C. baccatum se pretende avanzar en el conocimiento de variables de relevancia en la reproducción, en especial los efectos del ambiente.

Investigación de defectos genéticos en el sistema de la fosforilasa hepática en pacientes argentinos. Definición nosológica mediante una estrategia de análisis molecular. Investigation of genetic defects in the hepatic phosphorylase system in Argentinean patients. Nosological definition by molecular analysis strategy.

Las Enfermedades de Atesoramiento de Glucógeno (EAGs) también llamadas Glucogenosis comprenden un grupo de entidades causadas por una deficiencia enzimática específica relacionada con la vía de síntesis o degradación de esta macromolécula. La heterogeneidad fenotípica de los pacientes afectados dificulta la identificación de las diferentes variantes de EAG y por ende la correcta definición nosológica. En el Centro de Estudio de las Metabolopatías Congénitas, CEMECO, se fueron definiendo los diferentes tipos de Glucogenosis a través de una estrategia multidisciplinaria que integra distintos niveles de investigación clínica y complementaria, laboratorio metabólico especializado, enzimático, histomorfológico y de análisis molecular. Sin embargo, en algunos enfermos, entre los que se encuentran aquellos con defectos en el sistema de la fosforilasa hepática (EAG-VI y EAG-IX), la exacta definición nosológica aún no está resulta. La EAG-VI se refiere a un defecto en la fosforilasa hepática, enzima codificada por el gen PYGL, mientras que la EAG-IX es causada por un defecto genético en una de las subunidades de la fosforilasa b quinasa hepática codicadas por los genes PHKA2, PHKB y PHKG2, respectivamente. El objetivo del presente trabajo es propender a la definición nosológica de pacientes con defectos en el sistema de la fosforilasa mediante una estrategia de análisis molecular investigando los genes PYGL, PHKA2, PHKB y PHKG2. Los pacientes incluidos en este estudio deberán ser compatibles de padecer una EAG-VI o EAG-IX sobre la base de síntomas clínicos y hallazgos bioquímicos. La metodología incluirá la determinación de la enzima fosforilasa b quinasa en glóbulos rojos y dentro del análisis molecular la extracción de DNA genómico a partir de sangre entera para la amplificación por PCR de los exones más las uniones exon/intron de los genes PHKG2 y PYGL y la extracción de RNA total y obtención de cDNA para posterior amplificación de los cDNA PHKA2 y PHKB. Todos los fragmentos amplificados serán sometidos a análisis de secuencia de nucleótidos. Resultados esperados. Este trabajo, primero en Argentina, permitirá establecer las bases moleculares de los defectos del sistema de la fosforilasa hepática (EAG-VI y EAG-IX). El poder lograr este nivel de investigación traerá aparejado, una oferta integrativa en el vasto capítulo de las glucogenosis hepáticas, con extraordinaria significación en la práctica asistencial para el manejo, pronóstico y correspondiente asesoramiento genético. Hepatic glycogen storage diseases (GSDs) are a group of disorders produced by a deficiency in a specific protein involved in the metabolism of glycogen causing different types of GSDs. Phenotypic heterogeneity of affected patients difficult to identify the different GSD variants and therefore the correct definition of the disease. In the “Centro de Estudio de las Metabolopatías Congénitas”, CEMECO, were defined the different GSD types by a protocol which included complex gradual levels of clinical, biochemical, enzymatic and morphological investigation. However, in some patients, like those one with defects in the hepatic phosphorylase system (GSD-VI and GSD-IX) the exact definition of the disease has not yet been resolved. The GSD-VI is produced by a defect in the PYGL gen that encode the liver phosphorylase, while the GSD-IX is caused by a genetic defect in one of the Phosphorylase b kinase subunits, encoded by the PHKA2, PHKB and PHKG2 genes, respectively. The aim of the present study is to define the phosphorylase system defects in argentinian patients through a molecular strategy that involve the investigation of PYGL, PHKA2, PHKB and PHKG2 genes. Patients included in the present study must be compatible with a GSD-VI or GSD-IX on the bases of clinical symptoms and biochemical findings. The phosphorylase b kinase activity will be assay on in blood red cells. The molecular study will include genomic DNA extraction for the amplification of PHKG2 and PYGL genes and the total RNA extraction for amplification of the PHKA2 and PHKB cDNA by PCR. All PCR-amplified fragments will be subjected to direct nucleotide sequencing. This work, first in Argentina, will make possible to establish the molecular basis of the defects on the hepatic phosphorylase system (GSD-VI and GSD IX). To achieve this level of research will entail advance in the study of the hepatic glycogen storage disease, with extraordinary significance in the treatment, prognosis and the genetic counselling.

Estudio de la muerte celular por Apoptosis en el ovario de Columba maculosa y Columbina picui (Aves:Columbidae) durante el Ciclo Reproductivo Anual. Study of cell death by apoptosis in the ovary of Columba maculosa and Columbina picui (Aves: Columbidae) during the reproductive annual cycle.

En este trabajo se analizarán las características estructurales, cuantitativas y el proceso de muerte celular por apoptosis en el ovario de C. maculosa y C.picui con el objetivo de aportar conocimientos básicos a la biología reproductiva de estas aves. Cincuenta hembras adultas de cada especie se capturarán en el Dpto. Río Primero (Pcia. de Cba), R. Argentina, durante el ciclo reproductivo 2011-2012. Las muestras de ovarios se fijaran en Formalina Neutra pH 7.0, procesarán con la técnica de inclusión en parafina y colorearan con Hematoxilina /Eosina y Reacción Nuclear de Feulgen. Cinco muestras serán utilizadas para la determinación de muerte celular por apoptosis con la técnica de TUNEL. Se estudiarán las características morfohistológicas del ovario de C.maculosa y C. picui e identificarán, categorizarán y cuantificarán los folículos atrésicos no bursting (folículos atrésicos previtelogénicos y vcitelogénicos pequeños que conservan la integridad de la pared folicular) y bursting (folículos vitelogénicos mayores de 2 mm que liberar el contenido folicular por ruptura de la pared folicular) durante el ciclo reproductivo anual. Mediante la marcación de ADN fragmentado, se revelará la muerte celular por apoptosis en las células granulosas de los folículos atrésicos. Se compararán las semejanzas y diferencias estructurales y cuantitativas y el proceso de muerte celular en los folículos regresivos entre las dos especies. Los resultados de este trabajo representarán un importante aporte al conocimiento de la atresia folicular como así también a la muerte celular, un proceso estrechamente asociado a la misma, aún poco estudiado en el ovario de las aves. In this work we analyse the structural, quantitative and the process of the cell death by apoptosis in the ovary of Columba maculosa and Columbina picui in order of providing basic knowledge of reproductive biology of these birds. Fifty adult female of each species will be caught in the Río Primero (Pcia. Cba) R.Argentina, during 2011 - 2012. The ovarian samples will be fixed en neutral buffered Formalin (pH 7.0), processed with the technique of inclusion in paraffin and stained with Haematoxylin - Eosin, and nuclear Reaction of Feulgen. Five samples will be used to reveal cell death by apoptosis with the technique of TUNEL. Will be studied the morphohistological characteristics of ovary of both species, and identify, categorize and quantify the atretic follicles over the cycle, the non-bursting (pre-vitellogenic follicles and vitellogenic small < 2 mm, involution an without follicular rupture) and bursting (vitellogenic follicle > 2 mm in the which the yolk falls into the peritoneum due to rupture of with out put of the wall follicular). Will be revealed DNA fragmentation in the granulosa cells of the atretic follicles. Will compared the similaritues and differences in structure and quantitative and the cell death process by apoptosis in the regresive follicles, between the two species. The results of this study represent an important contribution to the knowledge of follicular atresia as well cell death a process closely associated with it and still poorly studied in the ovary of birds.

Estrategias reproductivas de lagartos del género tupinambis en distintos contextos ecológicos de la Provincia de Córdoba. Reproductive strategies of lizards of genus tupinambis in different ecological contexts of the Cordoba province.

Los caracteres de historia de vida son sensibles a la variación histórica o actual de los factores ambientales. Estudiar dicha variabilidad mediante la realización de estudios comparativos permite obtener evidencias sobre las causas de la evolución de ciertos caracteres. Los lagartos son excelentes modelos para el estudio de selección sexual y evolución del comportamiento social y reproductivo debido a que su relativa baja dispersión podría tener consecuencias evolutivas profundas en el desarrollo de distintas estrategias, ya que las poblaciones, al encontrarse más aisladas, podrían verse influenciadas por las fuerzas selectivas locales, mostrando una alta heterogeneidad espacial y temporal. Por eso nos propusimos realizar este trabajo para evaluar si existen diferentes estrategias reproductivas en los lagartos del género Tupinambis en distintos contextos ecológicos de la provincia de Córdoba. Para ello analizaremos distintas características de la historia de vida en poblaciones de estas especies tales como estructura de tamaño, sexo operativo, frecuencia reproductiva, tamaño de camada, condición corporal reproductiva, tamaño de madurez sexual, características espermáticas, elección de sitios de nidificación, etc. Además analizaremos la estructura genética de las poblaciones para inferir procesos demográficos históricos y patrones actuales de flujo génico y conectividad. The life history traits are sensitive to historical or current variation of environmental factors. Studying this variability by performing comparative studies allows obtaining evidence on the causes of the evolution of certain characters. Lizards are excellent models for studying sexual selection and evolution of social and reproductive behavior because their relatively low dispersal capabilities could have profound evolutionary consequences in the development of different strategies, since isolated populations may be stronger influenced by local selective forces, showing a high spatial and temporal heterogeneity. We decided to perform this study to assess whether there are different reproductive strategies in lizards of the genus Tupinambis in different ecological contexts of the Cordoba province. We will analyze different life history traits in populations of these species such as size structure, operational sex ratio, reproductive frequency, litter size, body condition, size at sexual maturity, sperm characteristics, choice of nesting sites, etc.. We also analyzed the genetic structure of populations to infer historical demographic processes and current patterns of gene flow and connectivity.

Age, growth and reproductive biology of whiting Merlangius merlangus (Linnaeus 1758) in the Celtic Sea

Age, growth and reproductive biology were investigated for whiting (Merlangius merlangus) captured from the Celtic Sea (ICES division Vllg), for the period January 2001 to January 2002. Females dominated the sex ratio of 1: 2.25. The relative abundance of females exceeded the number of males in all length classes. The relationship between weight (g) and total length (cm) was the same for male and female whiting. A total of 973 fish were aged and the maximum age recorded was 11 years. Results from an intercalibration exercise showed 87% agreement in age readings between the author and an expert in ageing whiting at the Marine Institute. Females were dominated by 2 year olds, while males were dominated by 3 year olds. Lx was estimated as 38cm and a growth rate was calculated K = 0.3769 year'1. Females were fully recruited to the fishery at 3 years of age, while the age at full recruitment (tr) for males was 4 years. Female whiting spawned from late February to June 2001 and matured at a total length of 23 cm in their first year. Female whiting reached L5o at a total length of 28 cm and 2.7 years of age. Male whiting spawned from February to June 2001. They matured at a total length of 21 cm and in their first year. Male whiting reached L50 at a total length of 30.4 cm and 3.6 years of age. The following critical points should be taken into account in the management of the Celtic Sea whiting stock: An Fpa should be established in order to assess the current level of fishing mortality; The maturity ogives need further study; The extent of gutting of large fish before landing by fishers in the fleet should be investigated and the apparent decline in size of 4 - 7 year old fish in the Celtic Sea between 1996 and 2001 needs to be assessed.