Probing the role of the fully conserved Cys126 in triosephosphate isomerase by site-specific mutagenesis - distal effects on dimer stability

Cys126 is a completely conserved residue in triosephosphate isomerase that is proximal to the active site but has been ascribed no specific role in catalysis. A previous study of the C126S and C126A mutants of yeast TIM reported substantial catalytic activity for the mutant enzymes, leading to the suggestion that this residue is implicated in folding and stability [Gonzalez-Mondragon E et al. (2004) Biochemistry43, 3255–3263]. We re-examined the role of Cys126 with the Plasmodium falciparum enzyme as a model. Five mutants, C126S, C126A, C126V, C126M, and C126T, were characterized. Crystal structures of the 3-phosphoglycolate-bound C126S mutant and the unliganded forms of the C126S and C126A mutants were determined at a resolution of 1.7–2.1 Å. Kinetic studies revealed an approximately five-fold drop in kcat for the C126S and C126A mutants, whereas an approximately 10-fold drop was observed for the other three mutants. At ambient temperature, the wild-type enzyme and all five mutants showed no concentration dependence of activity. At higher temperatures (> 40 °C), the mutants showed a significant concentration dependence, with a dramatic loss in activity below 15 μm. The mutants also had diminished thermal stability at low concentration, as monitored by far-UV CD. These results suggest that Cys126 contributes to the stability of the dimer interface through a network of interactions involving His95, Glu97, and Arg98, which form direct contacts across the dimer interface.

Determinación de la calidad de la carne bovina mediante inspección sanitaria (ISC) en el matadero PROINCASA

La carne bovina hoy en día se considera un producto de exportación y Nicaragua es el mayor exporta dor de este producto en Centroamérica, es así que la carne bovina en conjunto con el café son los productos de exportación que aportan los ingresos mas altos del país (Schütz et al., 2004). Conociéndose que el bovino en el matadero sufre una transformación a materia prima, donde los principales productos y subproductos obtenidos de la matanza son los cortes selectos, cortes industriales, víscera s comestibles y otros. Con base en la importancia que representa la producción de carne bovina para el país, se decidió realizar el presente estudio que tiene por titulo “Determinación de la Calidad de la Carne Bovina mediante Inspección Sanitaria (ISC) en el matadero PROINCASA”, con el objetivo de determinar la calidad de la carne bovina la cual se obtuvo mediante revisión de los registros del matadero para el año 2005 e Inspección Sanitaria de la Carne (ISC) directo para el primer semestre del año 2006, donde identificamos las patologías mas frecuentes en glándulas mamarias (ubres) con un 1.11% cueros 0.59% e hígados 0.50%. Así como también se lograron determinar perdidas económicas las cuales reflejan C$ 965 367.00 Córdobas. Con este trabajo de investigación se concluye que no se alcanza el aprovechamiento completo de los animales debido al alto porcentaje de afecciones patológicas que se diagnostican mediante Inspección Sanitaria de la Carne(ISC).

Evaluación de un sistema de riego por aspersión de efluentes de tambo

Resumen: Según los últimos datos publicados por SENASA, a marzo de 2011, el número de unidades productivas con tambo a nivel nacional fue de 11.646 (Informe Acrea, 2011), registrándose al mismo tiempo un aumento de la cantidad de vacas por tambo, en la producción individual y por consiguiente en la producción diaria por tambo (Castignani, 2009). El incremento de la cantidad de efluentes generados por la intensificación de los sistemas lecheros es hasta ahora un problema que puede traer aparejado efectos negativos en la contaminación del medio ambiente. Generalmente los productores ven a dichos efluentes como desperdicios derivados del proceso de ordeño, que se acumulan en fosas donde, es posible que rebalsen a cursos naturales de agua o comienzan acumularse en grandes lagunas en las proximidades de los tambos provocando malos olores y atracción de insectos. El manejo de estos residuos es determinante no solo para reducir la transferencia de nutrientes desde la pastura hacia los corrales, sino para limitar su efecto negativo sobre el ambiente, la salud humana y animal (Taverna et al, 2004). En siguiente trabajo experimental se realizó una descripción de los sistemas que existen en la actualidad para el manejo de los efluentes en las instalaciones lecheras. Además se hace una reseña de la legislación que existe en Argentina con respecto al uso y vertido de este tipo de efluentes. Se realizaron encuestas para evaluar el funcionamiento del aspersor de efluentes comercializado por la empresa Delaval Bosio. Los resultados muestran una tasa de aplicación de 53,16 l/m2/día, con una uniformidad de distribución promedio de 50 %. El equipo presentó algunos inconvenientes según lo expresado por sus usuarios, tales como el tapado de cañerías y la falta de atención técnica entre otros. Sin embargo, se reconoce al sistema como una alternativa de tratamiento de los efluentes generados, que de otra manera serían arrojados deliberadamente a cunetas, fosas y proximidades de las instalaciones. Con respecto a la calidad del efluente como abono cabe mencionar que con la tasa de aplicación expresada anteriormente los aportes de nutrientes serian de 288 kg/año de N y 114,76 kg/año de P respectivamente, representado de esta manera un ahorro para el productor en la incorporación de fertilizantes comerciales.

Power analysis for detecting trends in juvenile spotted seatrout abundance in Florida Bay

The spotted seatrout (Cynoscion nebulosus) is considered a key species relative to the implementation of the Comprehensive Everglades Restoration Plan (CERP). One of the goals of the CERP is to increase freshwater flows to Florida Bay. Increased freshwater flows can have potential positive and negative impacts on spotted seatrout populations. At low salinities, the planktonic eggs of spotted seatrout sink to the bottom and are not viable (Alshuth and Gilmore, 1994; Holt and Holt, 2002). On the other hand, increased freshwater flows can alleviate hypersaline conditions that could result in an expansion of the distribution of the early life stages of spotted seatrout (Thayer et al., 1999; Florida Department of Environmental Protection1). Thus it would be useful to develop a monitoring program that can detect changes in seatrout abundance on time scales short enough to be useful to resource managers. The NOAA Center for Coastal Fisheries and Habitat Research (NOAA) has made sporadic collections of juvenile seatrout using otter trawls since 1984 (see Powell et al, 2004). The results suggest that it might be useful to sample for seatrout in as many as eight different areas or basins (Figure 1): Bradley Key, Sandy Key, Johnson Key, Palm Key, Snake Bight, Central, Whipray and Crocodile Dragover. Unfortunately, logistical constraints are likely to limit the number of tows to about 40 per month over a period of six months each year. Inasmuch as few seatrout are caught in any given tow and the proportion of tows with zero seatrout is often high, it is important to determine how best to allocate this limited sampling effort among the various basins so that any trends in abundance may be detected with sufficient statistical confidence. (PDF contains 16 pages)

Field Guide to Nonindigenous Marine Fishes of Florida

The purpose of this field guide is to provide information on nonindigenous (i.e., non-native) fishes that have been observed in Florida’s marine waters. Introductions of non-native marine fishes into Florida’s waters could be intentional or unintentional, and are likely from a variety of sources, including aquarium releases, escape from aquaculture, loss due to extreme weather events (e.g., flooding from hurricanes), and possibly transfer with ballast water or hull-fouling. Presently the lionfishes (Pterois volitans and P. miles) are the only non-native marine fish species known to be established along the coast of Florida. All other marine fishes in this guide (except the euryhaline species, see below) have infrequent occurrences, occur singly or in small groups, and have not yet become self-sustaining populations. Aquarium releases are one of the major pathways whereby nonindigenous fishes gain access to new environments (Ruiz et al. 1997; Fuller et al. 1999). Most of the nonindigenous marine fishes found in Florida’s waters are thought to be aquarium fishes that either were illegally released into the ocean or escaped captivity (e.g., during severe storm/flooding events). Indeed, south Florida is a hotspot for nonindigenous marine aquarium fishes (Semmens et al. 2004). Increased public awareness of the problems caused by released or escaped aquarium fishes may aid in stemming the frequency of releases. For example, HabitattitudeTM (www.habitattitude.net) is a national public awareness and partnership campaign that encourages aquarists and water gardeners to prevent the release of unwanted aquarium plants, fish and other animals. It prompts hobbyists to adopt alternative actions when dealing with these aquatic plants and animals. (PDF file contains 133 pages.)

Stormwater runoff - modeling impacts of urbanization and climate change

Development pressure throughout the coastal areas of the United States continues to build, particularly in the southeast (Allen and Lu 2003, Crossett et al. 2004). It is well known that development alters watershed hydrology: as land becomes covered with surfaces impervious to rain, water is redirected from groundwater recharge and evapotranspiration to stormwater runoff, and as the area of impervious cover increases, so does the volume and rate of runoff (Schueler 1994, Corbett et al. 1997). Pollutants accumulate on impervious surfaces, and the increased runoff with urbanization is a leading cause of nonpoint source pollution (USEPA 2002). Sediment, chemicals, bacteria, viruses, and other pollutants are carried into receiving water bodies, resulting in degraded water quality (Holland et al. 2004, Sanger et al. 2008). (PDF contains 5 pages)

Looking for safe harbor in a crowded sea: Coastal space use conflict and marine renewable energy development

Technological advances in the marine renewable energy industry and increased clarity about the leasing and licensing process are fostering development proposals in both state and federal waters. The ocean is becoming more industrialized and competition among all marine space users is developing (Buck et al. 2004). More spatial competition can lead to conflict between ocean users themselves, and to tensions that spill over to include other stakeholders and the general public (McGrath 2004). Such conflict can wind up in litigation, which is costly and takes agency time and financial resources away from other priorities. As proposals for marine renewable energy developments are evaluated, too often decision-makers lack the tools and information to properly account for the cumulative effects and the tradeoffs associated with alternative human uses of the ocean. This paper highlights the nature of marine space conflicts associated with marine renewable energy literature highlights key issues for the growth of the marine renewable energy sector in the United States. (PDF contains 4 pages)

Climate change adaptation through coastal and use management: The context of environmental justice

Despite an increasing literary focus on climate change adaptation, the facilitation of this adaptation is occurring on a limited basis (Adger et al. 2007) .This limited basis is not necessarily due to inability; rather, a lack of comprehensive cost estimates of all options specifically hinders adaptation in vulnerable communities (Adger et al. 2007). Specifically the estimated cost of the climate change impact of sea-level rise is continually increasing due to both increasing rates and the resulting multiplicative impact of coastal erosion (Karl et al., 2009, Zhang et al., 2004) Based on the 2007 Intergovernmental Panel on Climate Change report, minority groups and small island nations have been identified within these vulnerable communities. Therefore the development of adaptation policies requires the engagement of these communities. State examples of sea-level rise adaptation through land use planning mechanisms such as land acquisition programs (New Jersey) and the establishment of rolling easements (Texas) are evidence that although obscured, adaptation opportunities are being acted upon (Easterling et al., 2004, Adger et al.2007). (PDF contains 4 pages)

Evolução geológica do Complexo Quirino, Terreno Paraíba do Sul, Setor Central da Faixa Ribeira, com base em dados isotópicos de Sm-Nd e Sr

A área estudada está inserida na Faixa Ribeira, Segmento Central da Província Mantiqueira (Almeida et al., 1973, 1977, 1981), que representa um cinturão de dobramentos e empurrões gerado no Neo-proterozóico/Cambriano, durante a Orogênese Brasiliana, na borda sul/sudeste do Cráton do São Francisco (Almeida, 1971, 1977; Cordani et al., 1967, 1973; Cordani & Brito Neves, 1982; Teixeira & Figueiredo, 1991). Neste contexto, o Complexo Quirino é o embasamento retrabalhado do Terreno Paraíba do Sul (Heilbron et al., 2004). O Complexo Quirino é formado por extensos corpos de ortognaisses foliados a homogêneos, leuco a mesocráticos, de granulometria média à grossa, composicionalmente variando entre granitóides tonal��ticos/granodioríticos a graníticos, e apresentando enclaves de rochas ultramáficas, máficas e cálcio-silicáticas (ricas em tremolita). Os ortognaisses tonal��ticos/granodioríticos apresentam porfiroblastos de plagioclásio e a hornblenda como máfico principal, contrastando com os de composição granítica que apresentam porfiroblastos de K-feldspato e biotita predominante. Como acessórios aparecem zircão, titanita, apatita e epidoto. Também estão associados a estes ortognaisses, granitóides neoproterozóicos que formam corpos individualizados ou lentes anatéticas no conjunto paleoproterozóico. Estes são compostos predominantemente por biotita gnaisse e hornblenda-biotita gnaisse. A análise litogeoquímicas dos ortognaisses do Complexo Quirino demonstrou a existência de duas séries magmáticas distintas. A primeira pertencente à série cálcio-alcalina de alto-K apresenta uma composição mais expandida granítica-adamelítica/granodioritica/tonal��tica e é correlacionável aos bt-ortognaisses e alguns hb-bt-ortognaisses. Os ortognaisses da série médio-K apresentam composição predominantemente tonal��tica, sendo correlacionáveis à maioria dos hornblenda-biotita gnaisses. Enclaves lenticulares de metapiroxeníticos e anfibolíticos ocorrem em muitos afloramentos. Também ocorrem granitóides neoproterozóicos de composição graníticas a quartzo-monzoníticas O estudo isotópico de Sm-Nd e Sr demonstrou que os ortognaisses da série cálcio-alcalina de alto-K e aqueles da série cálcio-alcalina de médio-K possuem idades modelo TDM variando entre paleoproterozóicas a arqueanas, consistentes com dados U-Pb em zircão publicados na literatura. A série cálcio-alcalina de alto-K é mais antiga (2308 9,2 Ma a 2185 8 Ma) do que a série calcio-alcalina de médio-K (2169 3 a 2136 14 Ma) e a existência de zircões herdados com idades mínimas de 2846 Ma e 2981 Ma para série de médio-K e 3388 16 para série de alto-K. Os granitóides brasilianos possuem idades de cristalização neoproterozóica correlacionada a Orogênese Brasiliana (602 a 627 Ma) (Viana, 2008; Valladares et al., 2002)./Com base nos dados de Sr e Sm-Nd foi possível caracterizar 4 grupos distintos. Os grupos 1 e 2 são formados por rochas de idade paleoproterozóica (2,1 a 2,3 Ga) com idades modelo TDM variando de 2,9 e 3,4 Ga, εNd entre -8,1 e -5,8 e 87Sr/86Sr(t) = 0,694707 (Grupo 1) e TDM variando de 2,5 a 2,7 Ga, εNd entre -5,8 e -3,1 e 87Sr/86Sr(t) = 0,680824 (Grupo 2), formados no paleoproterozóico com contribuição de uma crosta arqueana. O grupo 3 é formado por rochas juvenis de idade paleoproterozóica, com idades de cristalização variando entre 2,0 e 2,2 Ga e com idades modelo TDM variando de 2,1 a 2,2 Ga e εNd entre + 1,5 e + 1,2. O grupo 4 é formado durante o neoproterozóico (645 Ma) por rochas possivelmente de idade paleoproterozóico com idades modelo TDM igual a 1,7 Ga e εNd igual a -8,3.

Identificação do padrão biomecânico de condutas motoras em escolares: o caso da marcha

A constituição de uma base de dados normativa da marcha é essencial para o diagnóstico e o tratamento de padrões atípicos da locomoção (SUTHERLAND et al., 1997). Não obstante, são escassas as informações relativas aos padrões normais da marcha de crianças (GANLEY e POWERS, 2005), carência ainda mais evidente no tocante à produção acadêmica sobre o padrão biomecânico da locomoção da população de crianças brasileiras. Nesse sentido, especialistas em análise de marcha alertam para o fato que crianças de diferentes populações podem exibir diferentes padrões de marcha de acordo com os grupos étnicos das quais foram extraídas (MORENO-HERNÁNDEZ et al., 2010). Assim sendo, o objetivo do presente estudo foi descrever o padrão biomecânico da marcha de crianças hígidas brasileiras entre 6 e 11 anos de idade. Cento e vinte e duas crianças hígidas, entre seis e 11 anos de idade foram aleatoriamente recrutadas de um universo de 328 alunos. Os sujeitos foram alocados em três grupos etários: Grupo 1 (6-7 anos), Grupo 2 (8-9 anos) e Grupo 3 (10-11 anos). Para o registro das imagens das marchas das crianças foi utilizado um sistema de captura bidimensional de movimento a uma freqüência de aquisição de 30 Hz, composto por uma câmera Sony modelo HC 46 posicionada ortogonalmente a 6 metros da pista. Marcadores esféricos reflexivos de 20mm de diâmetro foram fixados em ambos os lados do corpo dos participantes. Os valores em bruto das coordenadas dos marcadores foram transformadas em coordenadas globais 2D (CALDWELL et al., 2004) e processadas no software SkillSpector (Versão 1.0). A estratégia de Hof (1996) foi utilizada para a normalização dos dados da marcha. Os comprimentos de passo e passada apresentaram uma tendência de aumento com o avanço da idade até 8-9 anos de idade, ao passo que a cadência dos passos apresentou uma tendência de diminuição até o mesmo período. Os números não-dimensionais não apresentaram qualquer tendência de alteração com o avanço da idade. Os três grupos etários apresentaram trajetórias angulares articulares semelhantes. O presente estudo constitui ação pioneira no que tange à descrição do padrão cinemático da marcha de crianças hígidas brasileiras entre 6 e 11 anos de idade. Assim sendo, consideramos que um primeiro passo foi dado no sentido da constituição de uma base de dados normativa da locomoção desses indivíduos.

Advantages of using crest nets to sample presettlement larvae of reef fishes in the Caribbean Sea

Identifying the spatial and temporal patterns of larval fish supply and settlement is a key step in understanding the connectivity of meta-populations (Sale et al., 2005). Because of the potentially dispersive nature of the pelagic larval phase of most reef fishes, tracking cohorts from hatching to settlement is extremely difficult (but see Jones et al., 1999). However, for many studies it is sufficient to sample larvae immediately before settlement. Many coral reef fish species use mangrove and seagrass beds as nursery habitats (Nagelkerken et al., 2001; Mumby et al., 2004) and larvae of these species must pass over the reef crest in order to arrive at their preferred settlement habitats. The ability to sample this new cohort of larval fishes provides opportunities for researchers to explore the intricacies of the transition from larva to juvenile (Searcy and Sponaugle, 2001). Quantifying the potential settlers also provides valuable information about the spatial and temporal supply of presettlement larvae (Victor, 1986). Therefore a number of larval sampling methods were developed, one of which is the use of crest nets (Dufour and Galzin, 1993).

Natural mortality rate, annual fecundity, and maturity at length for Greenland halibut (Reinhardtius hippoglossoides) from the northeastern Pacific Ocean

Mortality, fecundity, and size at maturity are important life history traits, and their interactions determine the evolution of life history strategies (Roff, 1992; Stearns, 1992; Charnov, 2002). These same traits are also important for population dynamics models (Hunter et al., 1992; Clark, 1999). It is increasingly important to accurately determine Greenland halibut (Reinhardtius hippoglossoides) life history traits and to correctly assess the status of its stocks because low recruitment or low biomass estimates have led to catch restrictions in the Bering Sea and Aleutian Islands (Ianelli et al.1), the Northeastern Arctic (Ådlandsvik et al., 2004), and the Northwest Atlantic (Bowering and Nedreaas, 2000).

Hatching date, nursery grounds, and early growth of juvenile walleye pollock (Theragra chalcogramma) off northern Japan*

Walleye pollock (Theragra chalcogramma) is widely distributed in the North Pacific Ocean and plays an important role in coastal subarctic ecosystems. The Japanese Pacific population of this species is one of the most important demersal fishes for commercial fisheries in northern Japan. The population is distributed along the Pacific coast of Hokkaido and the Tohoku area (Fig. 1), which is the southern limit of distribution of the species in the western North Pacific. In Funka Bay, the main spawning ground for this population, pollock spawn from December to March (Kendall and Nakatani, 1992). Planktonic eggs and larvae are transported into the bay, where juveniles usually remain until late July when they reach 60−85 mm in total length (Hayashi et al., 1968; Nakatani and Maeda, 1987). These juvenile pollock then migrate from Funka Bay eastward to the Doto area off southeastern Hokkaido (Honda et al., 2004). Many studies on eggs, larvae, and juveniles of the species have been conducted in or near Funka Bay, but little information is available on the ecology of the early life stages in the Tohoku area. Hashimoto and Ishito (1991) suggested that eggs are transported from Funka Bay southward to the Tohoku area by the coastal branch of the Oyashio Current, but there has been no study to verify this hypothesis.

Comparison of two approaches for estimating natural mortality based on longevity

Vetter (1988) noted that her review of the estimation of the instantaneous natural mortality rate (M) was initiated by a discussion among colleagues that identified M as the single most impor ta nt but least well-estimated parameter in fishery models. A lthough much has been accomplished in the inter vening years, M remains one of the most difficult parameters to estimate in fishery stock assessments. A number of novel approaches using tagging and telemetry data provide promise for making reliable direct estimates of M for a given stock (Hearn et al., 1998 ; Frusher and Hoenig, 2001; Hightower et al., 2001; Latour et al., 2003; Pollock et al., 2004). However, such methods are often impracticable and fishery scientists must approximate M by using estimates made for other stocks of the same or similar species or by predicting M from features of the species’ life history (Beverton and Holt, 1959; Beverton, 1963; Alverson and Carney, 1975; Pauly, 1980; Hoenig, 1983; Peterson and Wroblewski, 1984; Roff, 1984; Gunderson and Dygert, 1988; Chen and Watanabe, 1989; Charnov, 1993; Jensen, 1996; Lorenzen, 1996).

Protocols for conducting dolphin capture-release health assessment studies

Marine mammals, such as dolphins, can serve as key indicator species in coastal areas by reflecting the effects of natural and anthropogenic stressors. As such they are often considered sentinels of environmental and ecosystem health (Bossart 2006; Wells et al. 2004; Fair and Becker 2000). The bottlenose dolphin is an apex predator and a key component of many estuarine environments in the southeastern United States (Woodward-Clyde Consultants 1994; SCDNR 2005). Health assessments of dolphins are especially critical in areas where populations are depleted, show signs of epidemic disease and/or high mortality and/or where habitat is being altered or impacted by human activities. Recent assessments of environmental conditions in the Indian River Lagoon, Florida (IRL) and the estuarine waters surrounding Charleston, South Carolina (CHS) highlight the need for studies of the health of local bottlenose dolphins. While the condition of southeastern estuaries was rated as fair in the National Coastal Condition Report (U.S. EPA 2001), it was noted that the IRL was characterized by poorer than expected benthic communities, significant sediment toxicity and increased nutrient concentrations. Similarly, portions of the CHS estuary have sediment concentrations of aliphatic aromatic hydrocarbons, select inorganic metals, and some persistent pesticides far in excess of reported bioeffect levels (Hyland et al. 1998). Long-term trends in water quality monitoring and recent scientific research suggest that waste load assimilation, non-point source runoff impacts, contaminated sediments, and toxic pollutants are key issues in the CHS estuary system. Several ‘hot spots’ with high levels of heavy metals and organic compounds have been identified (Van Dolah et al. 2004). High concentrations of anthropogenic trace metals, polychlorinated biphenyls (PCB’s) and pesticides have been found in the sediments of Charleston Harbor, as well as the Ashley and Cooper Rivers (Long et al. 1998). Two superfund sites are located within the CHS estuary and the key contaminants of concern associated with these sites are: polycyclic aromatic hydrocarbons (PAH), lead, chromium, copper, arsenic, zinc and dioxin. Concerns related to the overall health of IRL dolphins and dermatologic disease observed in many dolphins in the area (Bossart et al. 2003) initiated an investigation of potential factors which may have impacted dolphin health. From May-August 2001, 35 bottlenose dolphins died in the IRL during an unusual mortality event (MMC 2003). Many of these dolphins were diagnosed with a variety of skin lesions including proliferative ulcerative dermatitis due to protozoa and fungi, dolphin pox and a vesicular dermatopathy of unknown etiology (Bossart et al. 2003). Multiple species from fish to dolphins in the IRL system have exhibited skin lesions of various known and unknown etiologies (Kane et al. 2000; Bossart et al. 2003; Reif et al. 2006). On-going photo-identification (photo-ID) studies have documented skin diseases in IRL dolphins (Mazzoil et al. 2005). In addition, up to 70% of green sea turtles in the IRL exhibit fibropapillomas, with the highest rates of occurrence being seen in turtles from the southern IRL (Hirama 2001).