分子生物光子学的进展

随着分子生物学和标记技术的进步，光学成像和分析有了很快的发展，无论是从理论上还是应用上都有所突破，展现了良好的前景。本文综述了近年来在分子生物探测中光子学的情况，简介了几种突破光学衍射极限的探测方法，着重阐明了与生化技术密切相关的荧光共振能量转移和荧光相关光谱显微术及其在分子生物学中的应用。

Polimorfismo genético de citocinas na população do Rio de Janeiro

Citocinas são moléculas que controlam e modulam a atividade de numerosas células por se ligarem a seus receptores específicos. As diferenças observadas na produção de citocinas entre indivíduos podem ser, pelo menos em parte, explicadas pelos polimorfismos genéticos como o polimorfismo de um único nucleotídeo (SNP). Em 181 indivíduos saudáveis não-aparentados da cidade do Rio de Janeiro (região Sudeste - Brasil), nós analisamos os polimorfismos de citocinas em genes que codificam para Fator de Necrose Tumoral-alfa (TNF-a), Fator de Crescimento Transformante-beta (TGF-b), Interleucina-10, Interleucina-6 e Interferon-gama (IFN-g). Reação em cadeia da polimerase utilizando-se iniciadores sequencia-específicos foi realizada com auxílio do kit comercial CytGen (One Lambda Inc. Canoga Park, CA, USA). Ao todo, 8 polimorfismos foram analisados: TNF-a (-308G/A); TGF-b (códon 10C/T, códon 25C/G); IL-10 (-1082A/G, -819T/C, -592A/C); IL-6 (-174C/G) e IFN-g (+874T/A). Os dados observados foram comparados a três grupos de população de diferentes regiões do Brasil (São Paulo, Paraná e Bahia) e a três populações de outros continentes (Itália, Eslováquia e Negros Norte-Americanos). O teste qui-quadrado foi utilizado para as comparações. Nossa análise da população do Rio de Janeiro mostrou que os as freqüências alélicas em IL-10, IL-6 e IFN-g são desigualmente distribuídos entre Brancos, Mulatos e Negros (p<0,05). A comparação com populações de outras regiões do Brasil revelou que Rio de Janeiro e Bahia possuem freqüências alélicas e genotípicas de TGF-b (códon 25) estatisticamente diferentes (p=0,004 e p=0,002, respectivamente). Ainda, a freqüência alélica na população do Rio de Janeiro é significativamente diferente quando comparada à população da Itália [IL-6 (-174), p=0,0092; e IFN-g (+874) p=0,0418)]; Eslováquia [IL-10(-1082), p=0,006; IL-6(-174), p=0,0002; e IFN-g(+874), p=0,0335]; e Afro-Americanos [IL-10(-819), p=0,0446; IL-6(-174), p<0,0001; e IFN-g(+874), p<0,0001]. Adicionalmente, observamos que a diferença na distribuição dos haplótipos em IL-10 (-1082/-819/-592) na população do Rio de Janeiro em comparação com a da Itália (p=0,0293) e Afro-Americanos (p=0,0025) é significativa. Portanto, concluímos que os polimorfismos em IL-10, IL-6 e IFN-g estão distribuídos de acordo com a etnia na população do Rio de Janeiro. A população do Rio de Janeiro possui freqüências de polimorfismos diferentes das populações de Bahia, Itália, Eslováquia e Afro-Americanos, mas semelhantes à população de São Paulo/Paraná. Nossas observações poderão ser úteis para futuros estudos e associação entre polimorfismos genéticos de citocinas e doenças na população do Rio de Janeiro.

Efeito da metformina no remodelamento miocárdico e renal em ratos obesos com resistência à insulina

Diversas evidências comprovam que a obesidade está associada a alterações estruturais e funcionais do coração em modelos humanos e animais. Outros estudos recentes também demonstram que a obesidade humana está associada com alterações na função e na estrutura vascular, especialmente em grandes e médias artérias. Estudos epidemiológicos têm confirmado que a obesidade é um fator de risco significativo para o aparecimento de proteinúria e de doença renal terminal em uma população normal. Com o objetivo de determinar as alterações morfológicas relacionadas ao remodelamento cardíaco, vascular e renal em um modelo experimental de obesidade induzida pelo glutamato monossódico (MSG) e os efeitos da metformina sobre estes achados, foram estudados 25 ratos divididos em cinco grupos: controle com 16 e 22 semanas (CON-16 e CON-22); obeso com 16 e 22 semanas (MSG-16 e MSG-22) e obeso + metformina (MET-22) 300mg/Kg/dia por via oral. A caracterização da resistência à insulina foi feita através da medida da insulina plasmática e cálculo do índice de HOMA-IR. As análises morfológicas e quantificação do colágeno miocárdico foram feitos pelo sistema de imagem Image Pro Plus analysis. A pressão arterial sistólica foi levemente maior no grupo MSG-22, adquirindo significância estatística quando comparada com o grupo MSG-16 (1222 vs 1082 mmHg, p<0,05). Por outro lado, o grupo MET-22 mostrou níveis mais baixos de pressão arterial (1181 mmHg), sem alcançar diferença significativa. No grupo de animais obesos, foi observado aumento na relação média-lumen com 16 semanas (39,93,7 vs 30,22,0 %, p<0,05) e com 22 semanas (39,81,3 vs 29,51,2%, p<0,05), que foi reduzida com o uso da metformina (31,50,9%). O depósito de colágeno na área perivascular no ventrículo esquerdo foi significativamente maior no grupo MSG-22 (1,390,06 vs 0,830,06 % no CON-22, p<0,01), sendo atenuado pela metformina (1,020,04%). No rim, a área seccional transversa das arteríolas intrarrenais foi semelhante entre os grupos (18,52,2 no CON-16; 19,93,7 no MSG-16; 18,93,1 no CON-22; 21,81,5 no MSG-22; 20,21,4 no MET-22). Foi observado aumento da área glomerular no grupo MSG-22 (141,34,5 vs 129,50,5 m2), mas sem significância estatística. Em conclusão, nos ratos com obesidade induzida pelo MSG, com resistência à insulina, as alterações cardíacas foram mais proeminentes do que as alterações renais. No coração foram observados sinais de remodelamento vascular hipertrófico nas pequenas artérias intramiocárdicas e evidências de fibrose miocárdica mais proeminente na área perivascular, alterações que foram, pelo menos parcialmente, atenuadas com o uso de metformina durante seis semanas, mostrando que esta droga pode ser benéfica na prevenção de complicações cardíacas, vasculares e renais associadas com a obesidade.

Age and growth of spiny dogfish (Squalus acanthias) in the Gulf of Alaska: analysis of alternative growth models

Ten growth models were fitted to age and growth data for spiny dogfish (Squalus acanthias) in the Gulf of Alaska. Previous studies of spiny dogfish growth have all fitted the t0 formulation of the von Bertalanffy model without examination of alternative models. Among the alternatives, we present a new two-phase von Bertalanffy growth model formulation with a logistically scaled k parameter and which estimates L0. A total of 1602 dogfish were aged from opportunistic collections with longline, rod and reel, set net, and trawling gear in the eastern and central Gulf of Alaska between 2004 and 2007. Ages were estimated from the median band count of three independent readings of the second dorsal spine plus the estimated number of worn bands for worn spines. Owing to a lack of small dogfish in the samples, lengths at age of small individuals were back-calculated from a subsample of 153 dogfish with unworn spines. The von Bertalanffy, two-parameter von Bertalanffy, two-phase von Bertalanffy, Gompertz, two-parameter Gompertz, and logistic models were fitted to length-at-age data for each sex separately, both with and without back-calculated lengths at age. The two-phase von Bertalanffy growth model produced the statistically best fit for both sexes of Gulf of Alaska spiny dogfish, resulting in L∞ = 87.2 and 102.5 cm and k= 0.106 and 0.058 for males and females, respectively.

Effective herding of flatfish by cables with minimal seafloor contact

Otter trawls are very effective at capturing flatfish, but they can affect the seaf loor ecosystems where they are used. Alaska f latf ish trawlers have very long cables (called sweeps) between doors and net to herd fish into the path of the trawl. These sweeps, which ride on and can disturb the seaf loor, account for most of the area affected by these trawls and hence a large proportion of the potential for damage to seaf loor organisms. We examined modifications to otter trawls, such that disk clusters were installed at 9-m intervals to raise trawl sweeps small distances above the seafloor, greatly reducing the area of direct seafloor contact. A critical consideration was whether flatfish would still be herded effectively by these sweeps. We compared conventional and modified sweeps using a twin trawl system and analyzed the volume and composition of the resulting catches. We tested sweeps raised 5, 7.5, and 10 cm and observed no significant losses of flatfish catch until sweeps were raised 10 cm, and those losses were relatively small (5–10%). No size composition changes were detected in the flatfish catches. Alaska pollock (Theragra chalcogramma) were captured at higher rates with two versions of the modified sweeps. Sonar observations of the sweeps in operation and the seaf loor after passage confirmed that the area of direct seafloor contact was greatly reduced by the modified sweep

Flatfish herding behavior in response to trawl sweeps: a comparison of diel responses to conventional sweeps and elevated sweeps

Commercial bottom trawls often have sweeps to herd fish into the net. Elevation of the sweeps off the seaf loor may reduce seafloor disturbance, but also reduce herding effectiveness. In both field and laboratory experiments, we examined the behavior of flatfish in response to sweeps. We tested the hypotheses that 1) sweeps are more effective at herding flatfish during the day than at night, when fish are unable to see approaching gear, and that 2) elevation of sweeps off the seafloor reduces herding during the day, but not at night. In sea trials, day catches were greater than night catches for four out of six flatfish species examined. The elevation of sweeps 10 cm significantly decreased catches during the day, but not at night. Laboratory experiments revealed northern rock sole (Lepidopsetta polyxystra) and Pacific halibut (Hippoglossus stenolepis) were more likely to be herded by the sweep in the light, whereas in the dark they tended to pass under or over the sweep. In the light, elevation of the sweep reduced herding, and more fish passed under the sweep. In contrast, in the dark, sweep elevation had little effect upon the number of fish that exhibited herding behavior. The results of both field and laboratory experiments were consistent with the premise that vision is the principle sensory input that controls fish behavior and orientation to trawl gear, and gear performance will differ between conditions where flatfish can see, in contrast to where they cannot see, the approaching gear.

Spatial and temporal variation in otolith chemistry for tautog (Tautoga onitis) in Narragansett Bay and Rhode Island coastal ponds

The elemental composition of otoliths may provide valuable information for establishing connectivity between fish nursery grounds and adult fish populations. Concentrations of Rb, Mg, Ca, Mn, Sr, Na, K, Sr, Pb, and Ba were determined by using solution-based inductively coupled plasma mass spectrometry in otoliths of young-of-the year tautog (Tautoga onitis) captured in nursery areas along the Rhode Island coast during two consecutive years. Stable oxygen (δ18O) and carbon (δ13C) isotopic ratios in young-of-the year otoliths were also analyzed with isotope ratio mass spectrometry. Chemical signatures differed significantly among the distinct nurseries within Narragansett Bay and the coastal ponds across years. Significant differences were also observed within nurseries from year to year. Classification accuracy to each of the five tautog nursery areas ranged from 85% to 92% across years. Because accurate classification of juvenile tautog nursery sites was achieved, otolith chemistry can potentially be used as a natural habitat tag.

Fish assemblages associated with three types of artificial reefs: density of assemblages and possible impacts on adjacent fish abundance

We evaluated the effectiveness of wooden artificial reefs (ARs) as fish habitat. Three types of ARs, made of cedar logs, broadleaf tree logs, and PVC pipes, respectively, were deployed in triplicate at 8-m depth off Maizuru, Kyoto Prefecture, Sea of Japan, in May 2004. Fish assemblages associated with each of the nine ARs were observed by using SCUBA twice a month for four years. Fish assemblages in the adjacent habitat were also monitored for two years before and four years after reef deployment. In the surveyed areas (ca. 10 m2) associated with each of the cedar, broadleaf, and PVC ARs, the average number of fish species was 4.14, 3.49, and 3.00, and the average number of individuals was 40.7, 27.9, and 20.3, respectively. The estimated biomass was also more greater when associated with the cedar ARs than with other ARs. Visual censuses of the habitat adjacent to the ARs revealed that the number of fish species and the density of individuals were not affected by the deployment of the ARs. Our results support the superiority of cedar as an AR material and indicate that deployment of wooden ARs causes no reduction of fish abundance in adjacent natural reefs.

Biomass and reproduction of Pacific sardine (Sardinops sagax) off the Pacific northwestern United States, 2003–2005

The Pacific sardine (Sardinops sagax) is distributed along the west coast of North America from Baja California to British Columbia. This article presents estimates of biomass, spawning biomass, and related biological parameters based on four trawl-ichthyoplankton surveys conducted during July 2003 –March 2005 off Oregon and Washington. The trawl-based biomass estimates, serving as relative abundance, were 198,600 t (coefficient of variation [CV] = 0.51) in July 2003, 20,100 t (0.8) in March 2004, 77,900 t (0.34) in July 2004, and 30,100 t (0.72) in March 2005 over an area close to 200,000 km2. The biomass estimates, high in July and low in March, are a strong indication of migration in and out of this area. Sardine spawn in July off the Pacific Northwest (PNW) coast and none of the sampled fish had spawned in March. The estimated spawning biomass for July 2003 and July 2004 was 39,184 t (0.57) and 84,120 t (0.93), respectively. The average active female sardine in the PNW spawned every 20–40 days compared to every 6–8 days off California. The spawning habitat was located in the southeastern area off the PNW coast, a shift from the northwest area off the PNW coast in the 1990s. Egg production in off the PNW for 2003–04 was lower than that off California and that in the 1990s. Because the biomass of Pacific sardine off the PNW appears to be supported heavily by migratory fish from California, the sustainability of the local PNW population relies on the stability of the population off California, and on local oceanographic conditions for local residence.

Seasonal variability in ichthyoplankton abundance and assemblage composition in the northern Gulf of Mexico off Alabama

Multiyear ichthyoplankton surveys used to monitor larval fish seasonality, abundance, and assemblage structure can provide early indicators of regional ecosystem changes. Numerous ichthyoplankton surveys have been conducted in the northern Gulf of Mexico, but few have had high levels of temporal resolution and sample replication. In this study, ichthyoplankton samples were collected monthly (October 2004–October 2006) at a single station off the coast of Alabama as part of a long-term biological survey. Four seasonal periods were identified from observed and historic water temperatures, including a relatively long (June–October) “summer” period (water temperature >26°C). Fish egg abundance, total larval abundance, and larval taxonomic diversity were significantly related to water temperature (but not salinity), with peaks in the spring, spring–summer, and summer periods, respectively. Larvae collected during the survey represented 58 different families, of which engraulids, sciaenids, carangids, and clupeids were the most prominent. The most abundant taxa collected were unidentified engraulids (50%), sand seatrout (Cynoscion arenarius, 7.5%), Atlantic bumper (Chloroscombrus chrysurus, 5.4%), Atlantic croaker (Micropogonias undulatus, 4.4%), Gulf menhaden (Brevoortia patronus, 3.8%), and unidentified gobiids (3.6%). Larval concentrations for dominant taxa were highly variable between years, but the timing of seasonal occurrence for these taxa was relatively consistent. Documented increases in sea surface temperature on the Alabama shelf may have various implications for larval fish dynamics, as indicated by the presence of tropical larval forms (e.g., fistularids, labrids, scarids, and acanthurids) in our ichthyoplankton collections and in recent juvenile surveys of Alabama and northern Gulf of Mexico seagrass habitats.

Observer-reported skate bycatch in the commercial groundfish fisheries of Alaska

We analyzed skate catch data collected by observers in the North Pacific Groundfish Observer Program (NPGOP) from 1998 through 2008 to document recent changes in the identification of skates by observers and to examine the species composition of observed skate catch in Alaska’s groundfish fisheries as well as recent trends in skate retention by commercial fishermen. Historically, almost all skate bycatch has been reported by NPGOP observers as “skate unidentified.” However, since 2004 observers have been trained to identify skates to the genus and species level. In 2008 over 95% of all skates were identified at least to the genus level, and over 50% were identified to species. The most common species of skates identified by observers in groundfish fisheries are Bathyraja parmifera (Alaska skate), Raja binoculata (big skate), and Bathyraja aleutica (Aleutian skate). Species composition of reported skate catch generally reflects recent survey-derived biomass estimates, with B. parmifera dominating the catches in the Bering Sea and, to a lesser extent, in the Aleutian Islands region, and species of the genus Raja dominating catches in the Gulf of Alaska. A relatively high percentage of the skate catch on longline vessels is still reported at the family or genus level because of difficulties in the identification of skates not brought onboard the vessel. For the larger skate species, the proportion retained for processing has increased in recent years as the market price for skate product has increased. Although observed skate catch does not give a complete account of skate bycatch in the fisheries of the region, observer data provide critical information for the appropriate management of skate populations in Alaska.

Effects of starvation on energy density of juvenile chum salmon (Oncorhynchus keta) captured in marine waters of Southeastern Alaska

We conducted laboratory starvation experiments on juvenile chum salmon (Oncorhynchus keta) captured in the neritic marine waters of northern Southeast Alaska in June and July 2003. Temporal changes in fish energy density (whole body energy content [WBEC], cal/g dry weight), percent moisture content, wet weight (g), length (mm), and size-related condition residuals were measured in the laboratory and were then compared to long-term field data. Laboratory water temperatures and salinities averaged 9°C and 32 psu in both months. Trends in response variables were similar for both experimental groups, although sampling intervals were limited in July because fewer fish were available (n= 54) than in June (n=101). Overall, for June (45-d experimental period, 9 intervals), WBEC, wet weight, and condition residuals decreased and percent moisture content increased, whereas fork length did not change. For July (20-d experimental period, 5 intervals), WBEC and condition residuals decreased, percent moisture content and fork length increased, and wet weight did not change. WBEC, percent moisture content, and condition residuals fell outside the norm of longterm data ranges within 10–15 days of starvation, and may be more useful than fork length and wet weight for detecting fish condition responses to suboptimal environments.

Accuracy of sex determination for northeastern Pacific Ocean thornyheads (Sebastolobus altivelis and S. alascanus)

Determining the sex of thornyheads (Sebastolobus alascanus and S. altivelis) can be difficult under field conditions. We assessed our ability to correctly assign sex in the field by comparing results from field observations to results obtained in the laboratory through both macroscopic and microscopic examination of gonads. Sex of longspine thornyheads was more difficult to determine than that of shortspine thornyheads and correct determination of sex was signif icantly related to size. By restricting the minimum size of thornyheads to 18 cm for macroscopic determination of sex we reduced the number of fish with misidentified sex by approximately 65%.

Measurement errors in body size of sea scallops (Placopecten magellanicus) and their effect on stock assessment models

Body-size measurement errors are usually ignored in stock assessments, but may be important when body-size data (e.g., from visual sur veys) are imprecise. We used experiments and models to quantify measurement errors and their effects on assessment models for sea scallops (Placopecten magellanicus). Errors in size data obscured modes from strong year classes and increased frequency and size of the largest and smallest sizes, potentially biasing growth, mortality, and biomass estimates. Modeling techniques for errors in age data proved useful for errors in size data. In terms of a goodness of model fit to the assessment data, it was more important to accommodate variance than bias. Models that accommodated size errors fitted size data substantially better. We recommend experimental quantification of errors along with a modeling approach that accommodates measurement errors because a direct algebraic approach was not robust and because error parameters were diff icult to estimate in our assessment model. The importance of measurement errors depends on many factors and should be evaluated on a case by case basis.