Utilização de sistema multiplex de marcadores do tipo INDELs em identificação humana por DNA

Os INDELs são polimorfismos de comprimento, gerados a partir de inserções e/ou deleções de um ou mais nucleotídeos. Os marcadores INDELs, que estão amplamente distribuídos pelo genoma e se caracterizam pela alta estabilidade devido à baixa taxa mutacional (10-9), podem ser analisados a partir da amplificação por PCR (Reação em Cadeia da Polimerase). A facilidade de análise e a possibilidade de construção de sistemas multiplex capazes de gerar amplicons curtos (menores que 100 pb) tornam os INDELs uma importante ferramenta para identificação humana por DNA. Para avaliar a eficiência e validar a metodologia que emprega os polimorfismos de inserção/deleção na identificação humana, utilizamos o sistema Indel-plex ID, capaz de amplificar simultaneamente 38 loci INDELs bialélicos de cromossomos autossomos. Diferentes amostras biológicas (cabelo, saliva, sangue, sêmen e urina) foram genotipadas apresentando reprodutibilidade entre todas as tipagens. A concentração mínima de DNA necessária para amplificação dos 38 loci INDELs foi de 0,5 ng. Artefatos do tipo split peaks foram observados em algumas amostras. Os produtos da PCR foram purificados em resina Sephadex proporcionando melhores condições de análise, redução de artefatos e aumento na intensidade média de fluorescência dos alelos amplificados. A eficiência do sistema Indel-plex ID na amplificação de DNA degradado foi verificada durante as análises das amostras de DNA extraídas de restos mortais (ossos e dentes). Comparativamente ao sistema Identifiler, o Indel-plex ID, se mostrou mais eficiente em termos de número de loci genotipados e qualidade de amplificação. Nas investigações de vínculos genéticos realizadas com o sistema Indel-plex ID foi possível corroborar resultados anteriores obtidos pela análise de marcadores STR. Nas análises com amostras in vivo foram obtidos valores máximos de Probabilidades de Paternidade de 99,99998%. Para casos envolvendo supostos pais falecidos, o sistema Indel-plex ID reforçou resultados obtidos com o sistema Identifiler e Minifiler. A Probabilidade de Paternidade de 99,953%, obtida com o sistema Indel-plex ID, conjugada com a Probabilidade de Paternidade de 99,957%, obtida como o sistema Minifiler, possibilitou um índice final de 99,99998%. Os resultados evidenciaram que os loci INDELs do sistema Indel-plex ID são altamente informativos, constituindo uma ferramenta importante em estudos de identificação humana e de relações de parentesco

Estudio del Sistema Renina Angiotensina como posible biomarcador para la mejora de los tratamientos de fecundación in vitro

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What Can the Brain Teach Us about Winemaking? An fMRI Study of Alcohol Level Preferences

Over the last few decades, wine makers have been producing wines with a higher alcohol content, assuming that they are more appreciated by consumers. To test this hypothesis, we used functional magnetic imaging to compare reactions of human subjects to different types of wine, focusing on brain regions critical for flavor processing and food reward. Participants were presented with carefully matched pairs of high- and low- alcohol content red wines, without informing them of any of the wine attributes. Contrary to expectation, significantly greater activation was found for low- alcohol than for high- alcohol content wines in brain regions that are sensitive to taste intensity, including the insula as well as the cerebellum. Wines were closely matched for all physical attributes except for alcohol content, thus we interpret the preferential response to the low- alcohol content wines as arising from top-down modulation due to the low alcohol content wines inducing greater attentional exploration of aromas and flavours. The findings raise intriguing possibilities for objectively testing hypotheses regarding methods of producing a highly complex product such as wine.

Freshwater pearl culture in India

Freshwater pearl culture is growing as a source of employment and income in many Southeast Asian countries. Bangladesh, Thailand, the Philippines and Vietnam have initiated freshwater pearl culture projects in recent years. In India, the Central Institute of Freshwater Aquaculture is carrying out research on identification of suitable local pearl mussel species (Genera Lamellidens)defining appropriate surgical implantation procedures, developing post-operative care procedures and captive pond culture of mussels.

Expression and Localization of Opioid Receptors in Male Germ Cells and the Implication for Mouse Spermatogenesis

The presence of endogenous opioid peptides in different testicular cell types has been extensively characterized and provides evidence for the participation of the opioid system in the regulation of testicular function. However, the exact role of the opioid system during the spermatogenesis has remained controversial since the presence of the mu-, delta-and kappa-opioid receptors in spermatogenic cells was yet to be demonstrated. Through a combination of quantitative real-time PCR, immunofluorescence, immunohistochemistry and flow cytometry approaches, we report for the first time the presence of active mu-, deltaand kappa-opioid receptors in mouse male germ cells. They show an exposition time-dependent response to opioid agonist, hence suggesting their active involvement in spermatogenesis. Our results contribute to understanding the role of the opioid receptors in the spermatogenesis and could help to develop new strategies to employ the opioid system as a biochemical tool for the diagnosis and treatment of male infertility.

Caribbean corals in crisis: record thermal stress, bleaching, and mortality in 2005

Background: The rising temperature of the world’s oceans has become a major threat to coral reefs globally as the severity and frequency of mass coral bleaching and mortality events increase. In 2005, high ocean temperatures in the tropical Atlantic and Caribbean resulted in the most severe bleaching event ever recorded in the basin. Methodology/Principal Findings: Satellite-based tools provided warnings for coral reef managers and scientists, guiding both the timing and location of researchers’ field observations as anomalously warm conditions developed and spread across the greater Caribbean region from June to October 2005. Field surveys of bleaching and mortality exceeded prior efforts in detail and extent, and provided a new standard for documenting the effects of bleaching and for testing nowcast and forecast products. Collaborators from 22 countries undertook the most comprehensive documentation of basin-scale bleaching to date and found that over 80% of corals bleached and over 40% died at many sites. The most severe bleaching coincided with waters nearest a western Atlantic warm pool that was centered off the northern end of the Lesser Antilles. Conclusions/Significance: Thermal stress during the 2005 event exceeded any observed from the Caribbean in the prior 20 years, and regionally-averaged temperatures were the warmest in over 150 years. Comparison of satellite data against field surveys demonstrated a significant predictive relationship between accumulated heat stress (measured using NOAA Coral Reef Watch’s Degree Heating Weeks) and bleaching intensity. This severe, widespread bleaching and mortality will undoubtedly have long-term consequences for reef ecosystems and suggests a troubled future for tropical marine ecosystems under a warming climate

Zooplankton population in the polluted environment of Thana Creek and Bombay Harbour

Hydrographical and biological parameters of Thana Creek and Bombay Harbour were studied to assess the prevailing water quality. Zooplankton samples were collected from various stations during January 1975 to July 1975. The qualitative distribution of zooplankton was found to be very irregular and fluctuating. Copepods were the dominant taxa followed by lucifers, chaetognaths, decapod larvae, ctenophores, hydromedusae, fish larvae and polychaetes. To a certain extent the distribution of zooplankton is affected by variation in salinity during different seasons, also along the length of the creek. Pronounced effect of pollution on zooplankton biomass was also observed.

Distribution of phytoplankton off Mithapur (Gujarat)

The area off Mithapur, Gujarat, India, is moderately rich in phytoplankton. The peak in phytoplankton population (av. 7.86 x 10 super(4)/l) was recorded in March. The average count of 0.71 x 10 super(4)/l observed in April was very low and later increased to 3.76 x 10 super(4)/l in December. Fifty-two species were recorded from the area. The most common species were Nitzschia closterium, N. pungens, N. seriata, N. sigma, Chaetoceros simplex, C. difficilis and C. sociale. The importance of Nitzschia spp as indicators of organically polluted environment was discussed.

Algae and water pollution in Mahi Estuary

Distribution of phytoplankton, chlorophyll A and phaeophytin was studied at different locations in the Mahi Estuary, Gujarat, India during 1982. The water quality at the discharge point was poor while the region away from it was relatively unpolluted. The results indicated a wide range of variation in phytoplankton population (7.68-5010, 96 x 10 super(4) cells/l, chl. a 2.22-58.22 mg/m super(3) and phaeophytin [0.20-10.21 mg/m super(3)]. The ratio of chl. a/phaeophytin were remarkably low at highly polluted stations. Higher abundance of the genera Oscillatoria and Nitzschia were recorded at polluted stations. The diversity of species at the unpolluted station was higher (1.56) as compared to the polluted station (1.07).

Phytoplankton distribution in the coastal waters off Porbandar (Saurashtra)

Phytoplankton population (3.37-56.09 x 10 super(4) cells/1) and pigments (chil. a 1.10-26.8 mg/m super(3)) in the waters off Porbandar indicated wide variations. Higher cell counts and lower diversity of species were encountered in the nearshore waters as compared to the waters of the seaward side. The concentration of dissolved oxygen was correlated with the phytoplankton population and pigments. 71 species belonging to 28 genera of algae were recorded. Species of Nitzschia were dominant almost at all stations. Species of Chaetoceros were very common along the middle and western transects.

Floating-body SOI memory: Concepts, physics and challenges

We present an overview of the single-transistor memory cells (lT-DRAMs), which are based on floating-body effects in SOI MOSFETs. The typical device architectures, principles of operation and key mechanisms for programming are described. The various approaches (Z-RAM, MSDRAM, etc) are compared in terms of performance and potential for aggressive scaling. ©The Electrochemical Society.

Physiological and biochemical study of over-ripened oocyte in the cultivated Caspian brown trout (Salmo trutta caspius) and determination of egg quality markers

To determine the best time for egg stripping after ovulation and over-ripened oocyte in the Caspian brown trout (Salmo trutta caspius), the eggs were retained in the parental abdominal cavity for 40 days post-ovulation (DPO) at 7±0.6°C. Eggs were stripped every 10-day interval in 4 treatment and were fertilized with a pool of semen obtained from 8 males. Also, the physiology and biochemistry of the eggs and ovarian fluids were studied. Results showed that the level of eyed eggs and hatched alevins declined with over-ripening time: that is, the expected amounts (90.65 ± 6.28% for eyeing and 86.33 ± 6.82% for hatching) in newly ovulated eggs (0–10 DPO) decreased to 0.67 ± 1.34% and 0.49 ± 0.98%, respectively, in over-ripened eggs (30–40 DPO). However, larval abnormalities remained constant for 30-days after ovulation. During the course of oocyte over-ripening, the pH of the ovarian fluid significantly decreased and the concentration of glucose, protein, calcium, iron, and aspartate aminotransferase activity significantly increased. Moreover, the concentration of protein, triglycerides, and aspartate aminotransferase activity in the eggs also changed. In the newly ovulated egg, the yolk consisted of homogenous tissue and its perivitelline space diameter had no considerable differences. With over-ripening, the yolk became heterogeneous, while chorion diameter and micropyle did not change. The perivitelline space diameter varied among different areas. The present study demonstrated that the best time to take Caspian brown trout eggs after ovulation at 7± 0.6°C was up to 10 DPO. Among the studied parameters of the egg and ovarian fluid, egg quality was related to both ovarian fluid parameters (e.g., pH, protein, aspartate aminotransferase, glucose, cholesterol, triglycerides, calcium, iron) and egg parameters (e.g., cholesterol, triglycerides, iron, aspartate aminotransferase). Thus, these parameters can be used as a egg quality markers in this species.

Cryopreservation of cynomolgus monkey (Macaca fascicularis) spermatozoa in a chemically defined extender

Aim: To establish a method for cynomolgus monkey sperm cryopreservation in a chemically defined extender. Methods: Semen samples were collected by electro-ejaculation from four sexually mature male cynomolgus monkeys. The spermatozoa were frozen in straws by liquid nitrogen vapor using egg-yolk-free Tes-Tris (mTTE) synthetic extender and glycerol as cryoprotectant. The effects of glycerol concentration (1%,3%, 5%, 10% and 15% [v/v]) and its equilibration time (10 min, 30 min, 60 min and 90 min) on post-thaw spermatozoa were examined by sperm motility and sperm head membrane integrity. Results: The post-thaw motility and head membrane integrity of spermatozoa were significantly higher (P < 0.05) for 5% glycerol (42.95 +/- 2.55 and 50.39 +/- 2.42, respectively) than those of the other groups (1%: 19.19 +/- 3.22 and 24.84 +/- 3.64; 3%: 34.23 +/- 3.43 and 41.37 +/- 3.42; 10%: 15.68 +/- 2.36 and 21.39 +/- 3.14; 15%: 7.47 +/- 1.44 and 12.90 +/- 2.18). The parameters for 30 min equilibration (42.95 2.55 and 50.39 2.42) were better (P < 0.05) than those of the other groups (10 min: 31.33 +/- 3.06 and 38. 98 +/- 3.31; 60 min: 32.49 +/- 3.86 and 40.01 +/- 4.18; 90 min: 31.16 +/- 3.66 and 38.30 +/- 3.78). Five percent glycerol and 30 min equilibration yielded the highest post-thaw sperm motility and head membrane integrity. Conclusion: Cynomolgus monkey spermatozoa can be successfully cryopreserved in a chemically defined extender, which is related to the concentration and the equilibration time of glycerol.

Ultrastructure of the Spermatozoa of the Yangtze Finless Porpoise (Neophocaena phocaenoides asiaeorientalis)

P>Semen sample was collected from two captive adult Yangtze finless porpoises (Neophocaena phocaenoides asiaeorientalis) during physical examination. One individual was aged about 9 years with body length 143 cm (total length) and body weight 46.1 kg in 2003. The age of the other was unknown and its body length was 147 cm and body weight was 43 kg in 2004. Ultrastructure of their spermatozoa was examined using scanning and transmission electron microscope. The sperm concentration was 4.17 x 10(9) spermatozoa per ml by the cytometer. The approximate dimensions of the spermatozoa were as follows: head length, 3.366 +/- 0.140 mu m (mean +/- SE, n = 15); head width, 1.896 +/- 0.099 mu m (n = 15); and neck length, 1.004 +/- 0.074 mu m (n = 10). The tail included midpiece, principal piece and terminal piece. The length of the midpiece was 1.882 +/- 0.077 mu m (n = 9). There is no apparent boundary between the principal piece and the terminal piece, so the length of the principal piece and the terminal piece was 44.612 +/- 3.485 mu m (n = 5). Total length of the spermatozoa was 53.314 +/- 4.580 mu m (n = 10). The acrosome covered approximately 45.8% of the anterior portion of the head.

单片机在定时数据采集器上的应用

本文介绍了一种自制单片机多通道数据采集及处理小系统。将其用在波浪能转换装置上，定时测量装置的发电量、电压、电流等。被测数据存放在带电保护的RAM内，系统配置RS232接口，与IBM微机通讯，便于采集的数据作进一步处理。