Search

As ectotherms, sea turtles are particularly sensitive to ambient temperature. Because of their charismatic megafauna and imperiled status, there is considerable interest as to how species in this taxon have responded to recent ocean warming and may respond to predicted warming trends. The fact that they are wide-ranging and evolutionarily ancient organisms suggests that marine tur- tles withstood changing climatic conditions in the past. To test if there are thermal cues that relate to dynamic nesting behaviours of loggerheads Caretta caretta and green turtles Chelonia mydas, we examined 20 yr (1989-2008) data sets of beach monitoring surveys and satellite-derived sea surface temperature (SST) from an important nesting site in east central Florida, USA, and adjacent Atlantic waters, respectively. For both species, median nesting dates became significantly earlier with higher May SSTs. However, the standard deviation of nest distributions, used as an analog for nesting sea- son length, decreased for loggerheads and, in contrast, increased for green turtles with a higher aver- age daily SST. This differential response between the species may reflect changes in reproductive physiology (e.g. internesting interval times and clutch numbers) and could have bearing on the future population dynamics of the 2 species.

Understanding the spatial movements of threatened marine species, such as sea turtles, is essential as a means of informing appropriate conservation management. Although novel tech- niques for tracking spatial movements are becoming more widely available (such as satellite track- ing), simple techniques such as mark-release-recapture remain effective. A flipper tagging and recovery program in Cuba tagged 210 loggerhead turtles over 14 yr and recovered 7% of the tags between 2 d and 3 yr later (mean = 296 d). All but one turtle was recaptured in Cuban waters, and data showed limited movement of turtles between northern and southern coasts. A further 50 turtles were recovered that had been tagged in foreign projects, the majority of which were from the USA (but also Mexico, The Bahamas, Canary Islands and Spain). A range of life stages of loggerhead tur- tles are found in Cuban waters year-round, and given that Cuba has the second largest reef in the Caribbean, it likely provides foraging habitat for significant numbers of loggerhead turtles from at least 6 different countries.

A generalized additive mixed modeling approach was used to assess somatic growth for juvenile green turtles Chelonia mydas at 4 sites in 3 ecologically distinct foraging habitats along the east central coast of Florida, USA. The 3 habitats were a man-made nuclear submarine turning basin (Trident Submarine Basin), an estuary (Indian River Lagoon), and oceanic sabellariid worm rock reefs (Sebastian Inlet and St. Lucie Power Plant). Turtles from the Indian River Lagoon site grew significantly faster than turtles from the Trident Submarine Basin and sabellariid worm rock reef sites. There were no significant differences in growth rates between the sabellariid worm rock reef and Trident Submarine Basin sites. Non-monotonic or dome-shaped growth rate functions reflecting an immature peak in growth rates were observed for all 3 habitats. Growth rates peaked in 1998 for turtles in the Trident Submarine Basin and sabellariid worm rock reef habitats; since then growth rates have declined. This temporal decline in growth rates may reflect density-dependent effects on growth as more juveniles recruit to Florida for aging grounds, a direct result of increases in nest production at the primary rookeries (Costa Rica, Florida and Mexico). Developmental habitats are important for the survival of juvenile marine turtles. This study illustrates the degree to which juvenile growth rates vary among developmental habitats, which ultimately can affect the rate of growth and recovery potential of nesting stocks.

Viral metagenomics, consisting of viral particle purification and shotgun sequencing, is a powerful technique for discovering viruses associated with diseases with no definitive etiology, viruses that share limited homology with known viruses, or viruses that are not culturable. Here we used viral metagenomics to examine viruses associated with sea turtle fibropapillomatosis (FP), a debilitating neoplastic disease affecting sea turtles worldwide. By means of purifying and shotgun sequencing the viral community directly from the fibropapilloma of a Florida green sea turtle, a novel single-stranded DNA virus, sea turtle tornovirus 1 (STTV1), was discovered. The single-stranded, circular genome of STTV1 was approximately 1,800 nucleotides in length. STTV1 has only weak amino acid level identities (25%) to chicken anemia virus in short regions of its genome; hence, STTV1 may represent the first member of a novel virus family. A total of 35 healthy turtles and 27 turtles with FP were tested for STTV1 using PCR, and only 2 turtles severely afflicted with FP were positive. The affected turtles were systemically infected with STTV1, since STTV1 was found in blood and all major organs. STTV1 exists as a quasispecies, with several genome variants identified in the fibropapilloma of each positive turtle, suggesting rapid evolution of this virus. The STTV1 variants were identical over the majority of their genomes but contained a hypervariable region with extensive divergence. This study demonstrates the potential of viral metagenomics for discovering novel viruses directly from animal tissue, which can enhance our understanding of viral evolution and diversity.

Marine turtle reproductive success is correlated with the stability and quality of the nesting environment. Female marine turtles show fidelity to nesting beaches, making artificial beach nourishment practices directly relevant to their recovery. We evaluated the impacts of artificial beach nourishment on Loggerhead (Caretta caretta) and Green turtles (Chelonia mydas) between artificially nourished and nonnourished beaches. We observed reduced nesting success (ratio of nesting emergences to emergences not resulting in nest deposition) for both species. This negative effect lasted for one season in Loggerheads and for at least one season in Green turtles. Physical attributes of the fill sand did not impede nesting attempts. We argue that the decrease in nesting success resulted from an altered beach profile not favorable for nest deposition, which subsequently improved in later seasons as the beach equilibrated to a more natural slope. We observed a 52.2% decrease in reproductive output (hatchlings km 21 yr 21 ) for Loggerheads one year postnourishment, with a 44.1% increase observed the two seasons postnourishment. In Green turtles, a 0.8% reduction was observed the first season postnourishment, despite a 13% increase in the nonnourished area. The reduction in reproductive output in both cases was primarily a consequence of decreased nesting success, lowering nest numbers. These results reveal stronger negative effects of beach nourishment on Loggerheads compared to Green turtles and the importance of minimizing excessive nonnesting emergences associated with artificial beach nourishment. Nourished areas also experienced more than 600% increase in the number of Loggerhead hatchlings disoriented by artificial lighting over two years postnourishment.

Chelonid fibropapillomatosis-associated herpesvirus (CFPHV) is an alphaherpesvirus believed to cause marine turtle fibropapillomatosis (FP). A serodiagnostic assay was developed for monitoring sea turtle populations for CFPHV exposure. CFPHV glycoprotein H (gH) expressed in recombinant baculovirus was used in an enzyme-linked immunosorbent assay (ELISA) to detect virus-specific 7S turtle antibodies. Using captive-reared green turtles ( Chelonia mydas ) with no history of virus exposure as “known negatives” and others with experimentally induced FP as “known positives,” the assay had 100% specificity but low sensitivity, as seroconversion was detected in only half of the turtles bearing experimentally induced tumors. Antibodies were detected only in samples collected after cutaneous fibropapillomas appeared, consistent with observations that tumors are significant sites of virion production and antigen expression and the possibility that prolonged/repeated virus shedding may be required for adequate stimulation of 7S antibody responses to gH. Natural routes of infection, however, may produce higher seroconversion rates. High gH antibody seroprevalences (∼80%) were found among wild green turtles in three Florida localities with different FP prevalences, including one site with no history of FP. In addition, all eight loggerhead turtles ( Caretta caretta ) tested were seropositive despite FP being uncommon in this species. The possibility that CFPHV infection may be common relative to disease suggests roles for environmental and host factors as modulators of disease expression. Alternatively, the possibility of other antigenically similar herpesviruses present in wild populations cannot be excluded, although antibody cross-reactivity with the lung/eye/trachea disease-associated herpesvirus was ruled out in this study.

The loggerhead sea turtle (Caretta caretta) is a federally threatened species and listed as endangered by the World Conservation Union (IUCN). We describe primers and polymerase chain reaction (PCR) conditions to amplify 11 novel tetranucleotide microsatellite loci from the loggerhead sea turtle. We tested primers using samples from 22 females that nested at Melbourne Beach, Florida (USA). Primer pairs yielded an average of 11.2 alleles per locus (range of 4–24), an average observed heterozygosity of 0.83 (range 0.59–0.96), and an average polymorphic information content of 0.80 (range 0.62–0.94). We also demonstrate the utility of these primers, in addition to primers for 15 loci previously described, for amplifying microsatellite loci in four additional species representing the two extant marine turtle families: olive ridley (Lepidochelys olivacea), hawksbill (Eretmochelys imbricata), green turtle (Chelonia mydas), and leatherback (Dermochelys coriacea).

Aim To critically review the status of the green sea turtle (Chelonia mydas) using the best available scientific studies as there is a prevailing view that this species is globally endangered and its marine ecosystem functions compromised. Location Ogasawara (Japan), Hawaii (USA), Great Barrier Reef (Australia), Florida (USA), Tortuguero (Costa Rica). Methods We compiled seasonal nesting activity data from all reliable continuous long-term studies (> 25 years), which comprised data series for six of the world's major green turtle rookeries. We estimated the underlying time-specific trend in these six rookery-specific nester or nest abundance series using a generalized smoothing spline regression approach. Results Estimated rates of nesting population increase ranged from c. 4-14% per annum over the past two to three decades. These rates varied considerably among the rookeries, reflecting the level of historical exploitation. Similar increases in nesting population were also evident for many other green turtle stocks that have been monitored for shorter durations than the long-term studies presented here. Main conclusions We show that six of the major green turtle nesting populations in the world have been increasing over the past two to three decades following protection from human hazards such as exploitation of eggs and turtles. This population recovery or rebound capacity is encouraging and suggests that the green turtle is not on the brink of global extinction even though some stocks have been seriously depleted and are still below historical abundance levels. This demonstrates that relatively simple conservation strategies can have a profound effect on the recovery of once-depleted green turtle stocks and presumably the restoration of their ecological function as major marine consumers.

We tracked 10 leatherback turtles by satellite from 2 Florida Atlantic Coast nesting beaches for a period ranging from 38 days to more than 454 days. Movement and foraging areas were often coastal, which contrasts with other satellite telemetry studies where leatherbacks are more pelagic. Using kernel home-range estimation we identified the primary internesting residence areas as well as Atlantic foraging areas. The primary internesting habitat was centered east–southeast of Cape Canaveral, Florida, from 2 to 60 km offshore and extending 215 km along the coast. Atlantic foraging areas were located primarily on the continental shelf from 30° to 50°N, and in an offshore area centered at 42°N, 65°W, as well as off Africa in the Mauritania upwelling. Seasonally, the location of these foraging areas changed, occurring on the North American continental shelf from March through November and off the shelf from December through February. One of the tracked turtles may have been killed with 17 other leathe...

We analyzed a 15-year (1989-2003) dataset of spatial nesting locations for Loggerhead and Green Turtles along a 40.5-km stretch of beach encompassing the Archie Carr National Wildlife Refuge along the Atlantic coast of Florida. To assess whether there are differences in spatial distribution influenced by temporal site-selection cues, we divided each season into quartiles and analyzed the autocorrelative patterns of the nest distributions within each time frame. Fundamen- tally, intraspecific differences in nest spatial patterns from the beginning to the end of the nesting season were minor. Though the temporal grain of the analyses may not be able to discern affects of fine-scale fluctuations (e.g., high- and low-tide events), these results suggest that environmental variables that change over the nesting season (e.g., ocean temperatures, daylength, and existing human activities) are not signifi- cantly influencing where these sea turtles place their nests.

Alterations in both vasculature and renin-angiotensin-aldoster-one system are a consistent finding in the metabolic syndrome. Adrenal tissue is highly vascularized and encounters blood flow, exceeding by far the volume expected for its size. Endothelial cells in the adrenal vasculature are therefore a major cellular component of adrenocortical tissue. The aim of the study was to analyze the cellular interaction between endothelial and steroid ' producing cells, focusing on endothelial cell-factor-mediated activation of aldosterone synthesis. The interaction between human endothelial (HUVECs) cell-conditioned medium and human adrenocortical (NCI-H295R) cells in vitro induced a significant surge in aldosterone secretion. The endothelial cell-conditioned medium together with angiotensin II and forskolin also potentiated aldosterone release by 1.5-fold and 2.6-fold, respectively, while preincubation of NCI-H295R cells for 24 h with endothelial cell-conditioned medium enhanced and sensitized the response of NCI-H295R to subsequent angiotensin II and forskolin stimuli by 2.5-fold and 2.2-fold, respectively. The increase in aldosterone release after preincubation with endothelial cell-conditioned medium was sensitive to cycloheximide and KN-93. Cellular conditioning with endothelial-cell factors exerts a hitherto unknown paracrine regulation of aldosterone production in human adrenocortical cells. This interaction may contribute to altered basal aldosterone release and have a role in patients with hypertension.

To study the movement patterns of Green Seaturtle (Chelonia mydas) populations in the Caribbean region using Cuban habitats, tag-recapture data from local (Cuban National Tagging Program 1989- 2002) and international programs (1959-2002) were compiled and compared. Of the 742 turtles tagged in Cuba at fishing areas, nesting beaches and head-start facilities, 5.5% were recaptured, mostly outside of Cuban waters and with a majority of these (76.9%) off the coast of Nicaragua. Green Seaturtles tagged elsewhere and recaptured in Cuba included head-started juveniles from Grand Cayman (45% of the total), Mexico (2.3%), and Florida (1.8%); wild juveniles from the Bahamas (14.1%), Bermuda (5.4%), and Florida (1.5%); and adults from Tortuguero (26%), Florida, USA (1.3%), Mexico (1%), Venezuela (1.3%), and U.S. Virgin Islands (0.3%). Recaptures of tags placed at sites north of Cuba (Bermuda and the Bahamas) clustered in the northeast region of Cuba, whereas those from the south (Grand Cayman) were recaptured in southern areas. Recaptures from Tortuguero tags were concentrated in the southeast and westward regions of Cuba. Turtles from the Bahamas, Grand Cayman, and Bermuda showed the highest recapture rates in Cuban habitats, with 3.2, 1.9, and 1.0% of the total number of tags applied, respectively. These results for a broad range of populations and across life stages underscore the regionwide significance of Cuban sites as critical habitats or migratory corridors.

Key words: Bayesian mixed-stock analysis, Indian River Lagoon, juvenile recruitment, loggerhead, marineturtlesIntroductionConservation of marine animals often is limited bythe ability of researchers to identify biologicaltrends and potential threats to organisms thatmake long distance migrations. For example, sal-mon production in California may be affected bylogging hundreds of miles inland (Caffereta andSpittler 1998) and marine turtle bycatch duringMediterranean shrimping operations decreasesbreeding populations on nesting beaches in Flor-ida and Mexico (Laurent et al. 1998). Attempts tosolve this problem by censusing rookeries, whilelogistically feasible, fail to incorporate predictivemodels and primarily record the results of trendsdisplayed by juveniles from distinct foraging areas(termed here as juvenile aggregations) in the priordecade. In the case of Florida marine turtles, theprimary measure of population change is thenumber of nests deposited on a beach, whichignores the effects of juvenile mortality caused bydisease (Work 2001), commercial fisheries (Crow-der 1995), and pollution on the future breedingpopulation. Understanding the factors dictatingjuvenile recruitment permits a more forwardlooking, predictive approach that incorporates theeffects of pollution, disease, natural disasters, andcommercial fisheries by-catch on juvenile popula-tions to predict future trends. Our detailed mixed-stock analysis of a large juvenile aggregation withseveral contributing rookeries reveals a novelapproach to understanding these factors and de-scribes patterns that are broadly applicable toregional and possibly global loggerhead rookeries.Loggerheads nest on sandy beaches throughouttemperate latitudes. The species was federally lis-ted as threatened in the North Atlantic in 1978 andis a CITES Appendix I listed species. The MarineTurtle Specialist Group and the IUCN Red Listconsider the loggerhead to be endangeredthroughout much of its range (Marine TurtleSpecialist Group 1996). Atlantic loggerheads leavetheir nesting beaches and enter oceanic currentsystems such as the Gulf Stream which laterbecomes part of the North Atlantic Gyre. Aftercircumnavigating the Atlantic for 3–10 years theyrecruit to a juvenile foraging area for the next 10–12 years (Carr 1986; Musick and Limpus 1997).Some of the juvenile (foraging) loggerheads in theNorth Atlantic recruit to juvenile foraging areas inthe Azores and Madeira (Bolten et al. 1998), butmany individuals enter foraging areas in IndianRiver Lagoon (IRL) on the east coast of centralFlorida.Aggregations of juvenile marine turtles mayinclude individuals from nesting beaches aroundthe globe, but until very recently, biologists hadnot devised a method for modeling contributionsof local rookeries to juvenile aggregations (Norr-gard and Graves 1995; Lahanas et al. 1998; Bass

During recent years our view of adipose tissue has been revolutionized. White adipose tissue (WAT) is no longer seen as mere energy store or provider of thermal and mechanical insulation. Neglect of WAT has been overcome by surprising discoveries in recent years, changing the view of this tissue towards a highly endocrine organ that is involved in a wide variety of physiological and pathophysiological processes. In this brief article we will focus on new developments in adipocyte and WAT biology. The appreciation of WAT as an endocrine organ will provide the basis for new and promising perspectives in the management of obesity and obesity-related diseases including diabetes, mellitus type II and arterial hypertension.

The onset of spring, noted by the timing of wildlife migratory and breeding behaviors, has been occurring earlier over the past few decades. Here, we examine 15 years of loggerhead sea turtle, Caretta caretta, nesting patterns along a 40.5 km beach on Florida’s Atlantic coast. This small section of beach is considered to be the most important nesting area for this threatened species in the western hemisphere. From 1989 to 2003, the annual number of nests fluctuated between 13 000 and 25 000 without a conspicuous trend; however, based on a regression analysis, the median nesting date became earlier by roughly 10 days. The Julian day of median nesting was significantly correlated with near-shore, May sea surface temperatures that warmed an average of 0.81C over this period. This marine example from warm temperate/subtropical waters represents another response of nature to recent climate trends.

The current study evaluates the interobserver reliability and intraobserver reproducibility of the Garden classification of femoral neck fractures, assesses the influence of a lateral radiograph on a fracture's classification, and determines the classification's impact on the surgeon's choice of operative treatment. Forty radiographs of femoral neck fractures were evaluated independently by five orthopaedic surgeons. Kappa values were calculated for interobserver reliability and intraobserver variability with respect to the readers' ability to assess the fractures using the Garden classification and to determine fracture displacement with and without access to a lateral radiograph. In 69% of the instances in which a reader changed the classification of a fracture, the proposed treatment of the fracture did not change. The Garden classification has poor interobserver reliability but good intraobserver reproducibility. The addition of a lateral radiograph does not seem to improve the reliability of the current Garden classification system but may improve the reader's ability to determine fracture displacement. To improve the reliability and usefulness of the Garden classification, the authors suggest that the classification should be modified to have only two stages (Garden A-nondisplaced or valgus impacted and Garden B-displaced) and to include the use of a lateral radiograph.

The Florida coastline from Melbourne Beach to Wabasso Beach is one the most important nesting areas for loggerhead turtles ( Caretta caretta ) in the Western Hemisphere and for green turtles ( Chelonia mydas ) in the United States. In this study, we quantified the spatial patterns of numerous loggerhead ( N ≈400,000) and green turtle ( N ≈14,000) and less numerous ( N ≈100) leatherback ( Dermochelys coriacea ) beach ascents from 1989–1999 in terms of their autocorrelative properties along 40.5 km within this critical reproductive zone. Nesting and non-nesting emergence patterns of loggerhead and green turtles were non-random, favoring the southern half of the study area. Perhaps due to low numbers or differences in nesting behavior, leatherback nest distributions were not significantly different from random. Loggerhead and green turtle nest locations exhibited similar clinal patterns. They were positively autocorrelated at distances less than 10 km and negatively at distances greater than 30 km. These patterns were significantly correlated interannually.

We describe isolation and characterization of 20 polymorphic tetranucleotide loci from the green turtle (Chelonia mydas). We identified an average of 12.5 alleles per locus based on screening of 31 individuals foraging in the Indian River Lagoon, Florida, USA. Observed heterozygosity ranged from 0.53 to 1.00, with a mean of 0.85. This microsatellite suite has a combined non-exclusion probability of identity of 2.45 × 10–28. These markers will complement those already available for green turtles in individual-based as well as population level genetic analyses.

Fibropapillomatosis is a neoplastic disease that is commonly found in the green turtles Chelonia mydas in tropical and subtropical regions of the world. In the current project, juvenile green turtles were captured with large-mesh tangle nets in the Indian River Lagoon and on nearshore reefs of Indian River County, Florida, USA, in 1998 and 1999. The purpose of the study was to determine the relationship between the severity of the disease and the general health of green turtles as indicated by blood parameters. All turtles were measured and examined, and the overall severity of the disease was rated by the size, number, and location of external fibropapilloma tumors. Hematocrit, total protein, and hemoglobin concentration were measured and compared with tumor scores (tumor severity appraisal). As the tumor score increased, the blood parameters of turtles decreased; for instance, the percentage of decrease in hematocrit for mildly afflicted, moderately afflicted, and severely afflicted groups were 2.6, 18.3, and 45.5%, respectively. Severely afflicted turtles suffered from anemia, while individuals with mild affliction did not.

Lung-eye-trachea disease-associated herpesvirus (LETV) is linked with morbidity and mortality in mariculture-reared green turtles, but its prevalence among and impact on wild marine turtle populations is unknown. An enzyme-linked immunosorbent assay (ELISA) was developed for detection of anti-LETV antibodies and could distinguish LETV-exposed green turtles from those with antibodies to fibropapillomatosis-associated herpesvirus (FPHV). Plasma from two captive-reared green turtles immunized with inactivated LETV served as positive controls. Plasma from 42 healthy captive-reared green turtles and plasma from 30 captive-reared green turtles with experimentally induced fibropapillomatosis (FP) and anti-FPHV antibodies had low ELISA values on LETV antigen. A survey of 19 wild green turtles with and 27 without FP (with and without anti-FPHV antibodies, respectively) identified individuals with antibodies to LETV regardless of their FP status. The seroprevalence of LETV infection was 13%. The presence of antibodies to LETV in plasma samples was confirmed by Western blot and immunohistochemical analyses. These results are the first to suggest that wild Florida green turtles are exposed to LETV or to an antigenically closely related herpesvirus(es) other than FPHV and that FPHV and LETV infections are most likely independent events. This is the first ELISA developed to detect antibodies for a specific herpesvirus infection of marine turtles. The specificity of this ELISA for LETV (ability to distinguish LETV from FPHV) makes it valuable for detecting exposure to this specific herpesvirus and enhances our ability to conduct seroepidemiological studies of these disease-associated agents in marine turtles.

A recently developed enzyme-linked immunosorbent assay (ELISA) was used to assess exposure of Florida wild green turtles Chelonia mydas to LETV, the herpesvirus associated with lung-eye-trachea disease (LETD). Plasma samples from 329 wild juvenile green turtles netted in the Indian River lagoon, along the Sebastian reef, or in the Trident basin (Indian River and Brevard Counties, Florida) were tested by ELISA for the presence of antibodies to LETV. Plasma samples from 180 wild juvenile green turtles were tested from these study sites to compare the prevalence of anti-LETV antibodies. While some plasma samples from each site contained anti-LETV antibodies (confirmed by Western blot analysis), plasma samples collected from the Indian River lagoon had statistically higher optical density values measured in the ELISA. No statistical differences were observed when these same plasma samples were analyzed for changes in the level of anti-LETV antibodies over 3 years (1997, 1998, and 1999). To explore the relationship between anti-LETV antibodies and fibropapillomatosis (FP), plasma from 133 green turtles scored for fibropapilloma tumor severity were tested by ELISA. There was no correlation between tumor severity and the presence of antibodies against LETV. Additional plasma samples collected from 16 tagged green turtles captured and sampled more than once (recaptures) were also tested to monitor antibody levels to LETV relative to the FP status of individual turtles over time. Again there was no clear relationship between FP tumor status and the presence of antibodies to LETV. Finally, ELISA tests on plasma from 13 nesting female turtles (9 green and 4 loggerhead) revealed high levels of anti-LETV antibodies in 11 individuals, including 2 loggerhead turtles. These results provide strong evidence that wild Florida green turtle populations at these 3 study sites are exposed to LETV or a closely related virus and that loggerhead turtles may be exposed as well. Based on a cutoff optical density value of 0.310, 71 out of the 329 wild Florida green turtles tested were seropositive for LETV antibodies (seroprevalence = 21.6%). In addition, no relationship between FP tumor severity or status and the presence of anti-LETV antibodies was found, further supporting the hypothesis that LETV and the FP-associated herpesvirus (FPHV) are separate infections of marine turtles.

Herpesviruses are associated with several diseases of marine turtles including lung-eye-trachea disease (LETD) and gray patch disease (GPD) of green turtles (Chelonia mydas) and fibropapillomatosis (FP) of green, loggerhead (Caretta caretta), and olive ridley turtles (Lepidochelys olivacea). The stability of chelonian herpesviruses in the marine environment, which may influence transmission, has not been previously studied. In these experiments, LETD-associated herpesvirus (LETV) was used as a model chelonian herpesvirus to test viral infectivity after exposure to seawater. The LETV virus preparations grown in terrapene heart (TH-1) cells were dialyzed for 24 to 120 hr against aerated artificial or natural seawater or Hank's balanced salt solution (HBBS). Fresh TH-1 cells were inoculated with dialyzed LETV, and on day 10 post-infection cells were scored for cytopathic effect. Virus samples dialyzed up to 120 hr were positive for the herpesvirus DNA polymerase gene by polymerase chain reaction. Electron microscopy revealed intact LETV nucleocapsids after exposure of LETV to artificial seawater or HBSS for 24 hr at 23 C. LETV preparations remained infectious as long as 120 hr in natural and artificial seawater at 23 C. Similar results were obtained with a second culturable chelonian herpesvirus, HV2245. LETV infectivity could not be detected after 48 hr exposure to artificial seawater at 30 C. Since LETV and HV2245 remain infectious for extended periods of time in the marine environment, it is possible that FP-associated and GPD-associated herpesviruses also may be stable. These findings are significant both for researchers studying the epidemiological association of herpesviruses with diseases of marine turtles and for individuals who handle turtles in marine turtle conservation efforts.

Endothelin-1 (ET-1) is a vasoconstrictive peptide which may play a relevant role in the pathogenesis of pulmonary hypertension (PH) in COPD patients. We assessed the correlations of plasmatic ET-1 levels with pulmonary function data, arterial blood gases and pulmonary haemodynamics in 21 COPD patients with moderate to severe airway obstruction (FEV1: 1.19+/-0.49 l, mean+/-SD). There were 11 hypoxemic patients (PaO265 mmHg). Six patients had resting PH (mean pulmonary artery pressure/=20 mmHg). Eight patients had exercising PH (PAP/=30 mmHg) during a short (6 min) 30 W steady state exercise. At rest, arterial ET-1 levels were significantly higher in COPD patients with hypoxemia when compared to those without hypoxemia (16.6+/-2.7 vs. 12.5+/-3.9 pM/l, P=0.02) and in COPD patients with PH when compared to those without PH (16.5+/-3.4 vs. 13+/-3.9 pM/l, P=0.04). Resting arterial ET-1 levels were negatively correlated with PaO2 (r=-0.45, P=0.05). At rest, the differences between mixed venous and arterial ET-1 levels were positively correlated with FEV1 (r=0.54, P=0.024). At exercise, the mean arterial ET-1 level was not significantly different from the mean resting ET-1 level (13.8+/-3.4 vs. 13.3+/-4 p M/l, NS). There were eight COPD patients who had decreasing arterial ET-1 levels between rest and the end of a 6 min exercise, and six COPD patients who had increasing arterial ET-1 levels. These variations of arterial systemic ET-1 levels from rest to exercise were negatively correlated with FEV1 (r=-0.66, P=0.01). We conclude that in COPD patients (1) at rest, arterial ET-1 levels are increased in hypoxemia or pulmonary hypertension and (2) at rest or during exercise, the turn-over of ET-1 may be dependent of the degree of the bronchial obstruction.

Recent data suggest that adrenocortical cells under pathological as well as under physiological conditions show neuroendocrine properties. Within the normal adrenal, this neuroendocrine differentiation seems to be restricted to cells of the zona glomerulosa and might be important for an autocrine regulation of adrenocortical function. In addition, such neuroendocrine differentiation is a common phenomenon in adrenocortical carcinomas and is therefore of clinical importance. In our studies, the expression of neuronal cell adhesion molecule (NCAM) could be shown in the zona glomerulosa of the normal human adrenal and in the human adrenocortical cell line NCI-H295 that also produces synaptophysin, a synaptic vesicle associated protein. In this chapter, data on neuroendocrine characteristics of adrenocortical cells are summarized and discussed.

Interleukin (IL)-6 is a potent activator of the human hypothalamic-pituitary-adrenal axis. After chronic administration of IL-6 in humans, there is a substantial elevation of cortisol, whereas ACTH levels are blunted. Thus, we investigated whether IL-6 and/or the IL-6 receptor (IL-6R) are expressed in the human adrenal gland and whether IL-6 could cause the release of steroid hormones by a direct action on adrenal cells in primary culture. The expression of IL-6 and IL-6R was investigated with RT-PCR and immunohistochemistry, and the effects on human adrenal steroidogenesis were tested with IL-6 in vitro. To avoid effects mediated by macrophages, we depleted adrenal primary cultures from macrophages using specific mouse antihuman CD68 and sheep antimouse IgG conjugated magnetic beads. The results showed that: 1) IL-6 and IL-6R are expressed in adrenal cell cultures, including all cell types and those depleted of macrophages; 2) IL-6R is mainly expressed in the zona reticularis and the inner zona fasciculata; positive signals from the zona glomerulosa and the medulla occurred in single cells; and 3) IL-6 regulates adrenal synthesis of mineralocorticoids, glucocorticoids, and androgens in vitro, dependent on time and dose, in the absence of macrophages. After 24 h, aldosterone secretion increased to 172 ± 28% sem, cortisol to 177 ± 27% sem, and dehydroepiandrosterone to 153 ± 20% sem of basal secretion. These findings, in combination with previous investigations, suggest that IL-6 exerts its acute action via the hypothalamus and the pituitary. In the adrenal gland, however, IL-6 seems to be a long-term regulator of stress response, integrating the responses of all cortical zones to stimuli from the immune and endocrine system.

Within the last years it has become evident that besides the hypothalamo-pituitary-adrenal axis, extrapituitary mechanisms exist that regulate the activity of the adrenal cortex. In this context, intra-adrenal regulatory mechanisms play an important role. Several secretory products from adrenomedullary cells are able to influence adrenocortical steroidogenesis. Since the main blood flow within the adrenal is directed centripetally from the cortex to the medulla, chromaffin cells should act on cortical cells in a paracrine manner. The morphological prerequisite for this regulatory pathway is seen in the close apposition of the two tissues. Within the mammalian adrenal, the two endocrine tissues are interwoven to an astonishing degree with cortical cells located within the medulla and vice versa. It is concluded from morphological and functional studies that paracrine interactions between cortex and medulla play an important role in the regulation of adrenocortical steroidogenesis. Microsc. Res. Tech. 36:520–533, 1997. © 1997 Wiley-Liss, Inc.

It is well established now that adrenocortical function, besides being regulated through systemic factors, is influenced by intra-adrenal mechanisms. In this context paracrine influences between the sympathoadrenal system and the adrenal cortex play an important role. As a prerequisite for these interactions, adrenal medullary cells and cortical cells are highly interwoven as revealed by immunohistochemistry. The potential role of VIP in the regulation of human adrenal steroidogenesis was now investigated in human adrenal cells in primary culture. The primary cultures contained both, cortical and chromaffin cells which were found to be in close cellular contact as revealed by immunocytochemistry. VIP enhanced cortisol secretion from adrenal cells in a dose-dependent manner with a maximal effect at 10(-7) M. VIP stimulated the release of dehydroepiandrosterone (DHEA), testosterone, androstenedione, and aldosterone significantly. The addition of propranolol, a beta-adrenergic antagonist, to the incubation medium attenuated VIP-induced corticosteroid secretion. It is concluded that VIP is a paracrine messenger in the human adrenal that could regulate adrenocortical function at least in part via catecholamines released from the medulla.

Sympathoadrenal regulation of adrenocortical steroidogenesis was studied on a physiological, cellular, molecular and morphological level. The effects of nerve activation and of epinephrine (EPI) on adrenal corticosteroid release were compared in isolated perfused pig adrenals with preserved nerve supply. Splanchnic nerve activation as well as perfusion with EPI provoked a significant release of cortisol, aldosterone and androstenedione. In cultured bovine adrenocortical cells steroid secretion and accumulation of P450scc, P450(17) alpha, P450c21 and P450(11) beta mRNAs were studied after stimulation with EPI with or without propranolol or phentolamine. Incubation with EPI stimulated steroidogenesis and increased the levels of all four P450-mRNAs. The beta-adrenergic antagonist propranolol totally blocked the effects of EPI while the alpha-antagonist phentolamine had no effect. Using immunohistochemistry, adrenals were studied morphologically. The contact zones of the two cell types were investigated on an electron microscopical level. Cortical and medullary cells were closely interwoven with cortical and chromaffin cells in direct apposition, providing the possibility for paracrine interactions. It is concluded that the release of corticosteroids can be stimulated through the sympatho-adrenal system. The stimulatory action of EPI upon adrenal steroid formation and accumulation of all four P450-mRNAs requires beta-adrenergic receptors. Taking into consideration the close colocalization of cortical and medullary tissue, this stimulation may be mediated by chromaffin cells in a paracrine manner.

Chromaffin cells probably are the most intensively studied of the neural crest derivates. They are closely related to the nervous system, share with neurons some fundamental mechanisms and thus were the ideal model to study the basic mechanisms of neurobiology for many years. The lessons we have learned from chromaffin cell biology as a peripheral model for the brain and brain diseases pertain more than ever to the cutting edge research in neurobiology. Here, we highlight how studying this cell model can help unravel the basic mechanisms of cell renewal and regeneration both in the central nervous system (CNS) and neuroendocrine tissue and also can help in designing new strategies for regenerative therapies of the CNS.

There is increasing evidence for an immune-adrenal interaction in which macrophages may play an important role. However, few data are available with respect to a human intra-adrenal macrophage system. In this study, we have investigated the density, distribution and phenotype of human adrenal macrophages using monoclonal antibodies. Macrophages are localized in all zones of the adrenal gland. These cells exhibit the phenotype of the phagocytotic macrophage compartment (CD11c+, KiM8+). At the ultrastructural level, macrophages are frequently attached to the endothelial wall, but also lie in direct contact with cortical and chromaffin cells. This investigation reveals the cellular basis for the possible role of macrophages in the local immune-neuroendocrine axis.

Ozobranchus margoi and Ozobranchus branchiatus are the only two species of marine turtle leeches (Ozobranchus spp.) known to inhabit the Atlantic coast of the United States and the Gulf of Mexico. In early reports of fibropapillomatosis (FP) in green turtles (Chelonia mydas), O. branchiatus was implicated as a vector in the transmission of Fibropapilloma-associated turtle herpesvirus (FPTHV). It is imperative that the leech species be identified to elucidate the role Ozobranchus spp. may play in disease transmission. In this study, Ozobranchus branchiatus has been identified for the first time on a loggerhead (Caretta caretta) turtle, and the molecular data for this species is now available for the first time in GenBank. Both species of leeches were also found infecting a single C. mydas. Using morphological taxonomy combined with distance- and character-based genetic sequence analyses, this study has established a DNA barcode for both species of Ozobranchus spp. leech and has shown it can be applied successfully to the identification of leeches at earlier stages of development when morphological taxonomy cannot be employed. The results suggest a different haplotype may exist for O. branchiatus leeches found on C. caretta versus C. mydas. Leech cocoon residue collected from a C. mydas was identified using the new method.

Our objective was to determine the effect of arachidonylethanolamide (anandamide, AEA) injected intracerebroventricularly (icv) into the lateral ventricle of the rat brain on submandibular gland (SMG) salivary secretion. Parasympathetic decentralization (PSD) produced by cutting the chorda tympani nerve strongly inhibited methacholine (MC)-induced salivary secretion while sympathetic denervation (SD) produced by removing the superior cervical ganglia reduced it slightly. Also, AEA (50 ng/5 μL, icv) significantly decreased MC-induced salivary secretion in intact rats (MC 1 μg/kg: control (C), 5.3 ± 0.6 vs AEA, 2.7 ± 0.6 mg; MC 3 μg/kg: C, 17.6 ± 1.0 vs AEA, 8.7 ± 0.9 mg; MC 10 μg/kg: C, 37.4 ± 1.2 vs AEA, 22.9 ± 2.6 mg). However, AEA did not alter the significantly reduced salivary secretion in rats with PSD, but decreased the slightly reduced salivary secretion in rats with SD (MC 1 μg/kg: C, 3.8 ± 0.8 vs AEA, 1.4 ± 0.6 mg; MC 3 μg/kg: C, 14.7 ± 2.4 vs AEA, 6.9 ± 1.2 mg; P < 0.05; MC 10 μg/kg: C, 39.5 ± 1.0 vs AEA, 22.3 ± 0.5 mg; P < 0.001). We showed that the inhibitory effect of AEA is mediated by cannabinoid type 1 CB1 receptors and involves GABAergic neurotransmission, since it was blocked by previous injection of the CB1 receptor antagonist AM251 (500 ng/5 μL, icv) or of the GABAA receptor antagonist, bicuculline (25 ng/ 5 μL, icv). Our results suggest that parasympathetic neurotransmission from the central nervous system to the SMG can be inhibited by endocannabinoid and GABAergic systems. Fil: Fernández Solari, José Javier. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Prestifilippo, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires; Argentina Fil: Vissio, Paula Gabriela. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ehrhart Bornstein, M.. Technische Universität Dresden; Alemania Fil: Bornstein, S. R.. Technische Universität Dresden; Alemania Fil: Rettori, V.. Universidad de Buenos Aires; Argentina Fil: Elverdín, Juan Carlos. Universidad de Buenos Aires; Argentina

We found a high concentration of galanin in extracts of porcine adrenal glands (114 pmol/g). By immunohistochemistry, galanin was localized to groups of medullary cells previously shown to produce norepinephrine. To study mechanisms for the release of galanin, we developed the following in vitro model: isolated perfused porcine adrenals with intact splanchnic nerve supply. When the nerves were electrically stimulated, epinephrine and norepinephrine secretion increased 276- and 291-fold, respectively, and galanin release increased up to 1,300-fold. Acetylcholine at 10(-6) M stimulated galanin release, and hexamethonium almost abolished the response to nerve stimulation. Galanin infusions had no effect on epinephrine and norepinephrine secretion in concentrations of 10(-8) and 10(-7) M, but increased both cortisol and aldosterone secretion (P less than 0.05). Splanchnic nerve stimulation in anesthetized pigs increased the concentration of galanin in the caval vein but not in arterial plasma. It is concluded that galanin, coreleased with catecholamines from the adrenal glands, may have endocrine functions but that galanin may also have local regulatory functions in the adrenals.

Recently, it has become evident that the adrenals play a key role in obesity as well as in the metabolic syndrome and their complications. On the one hand, adrenal steroids are involved in physiological regulation of adipose tissue and energy homeostasis and in the pathogenesis of cardiometabolic complications. On the other hand, fat cell-derived factors, adipocytokines, and lipids released from adipose tissue are involved in the modulation of adrenal steroidogenesis. Aldosterone plasma levels are elevated in obesity and in patients with the metabolic syndrome. Recent research has provided evidence that adipocytes secrete factors that stimulate adrenal mineralocorticoid release and sensitize the adrenal cortex to angiotensin II.

Sea turtle nesting presents a potential pathway to subsidize nutrient-poor dune ecosystems, which provide the nesting habitat for sea turtles. To assess whether this positive feedback between dune plants and turtle nests exists, we measured N concentration and delta15N values in dune soils, leaves from a common dune plant (sea oats [Uniola paniculata]), and addled eggs of loggerhead (Caretta caretta) and green turtles (Chelonia mydas) across a nesting gradient (200-1050 nests/km) along a 40.5-km stretch of beach in east central Florida, USA. The delta15N levels were higher in loggerhead than green turtle eggs, denoting the higher trophic level of loggerhead turtles. Soil N concentration and delta15N values were both positively correlated to turtle nest density. Sea oat leaf tissue delta15N was also positively correlated to nest density, indicating an increased use of augmented marine-based nutrient sources. Foliar N concentration was correlated with delta15N, suggesting that increased nutrient availability from this biogenic vector may enhance the vigor of dune vegetation, promoting dune stabilization and preserving sea turtle nesting habitat.

Three events of significance to the U. S. Transuranium and Uranium Registries (USTUR) occurred during this reporting period: 1. The search for a new Associate Director was successful in that Dr. Anthony C. (Tony) James was appointed to the position, 2. A five-year grant for the operation of the USTUR was approved by the U. S. Department of Energy; the previous grant cycles were for three years, 3. I retired from the USTUR Directorship on July 1, 2005 and Tony James became the new Director.

The development of the human fetal adrenals starts in the 6th week gestational age and adrenal C19 steroid production becomes of major importance for the maintenance of the pregnancy. Therefore, in the present study, human fetal adrenals at 6 weeks of gestational age were immunostained for 17 alpha-hydroxylase, the key enzyme for the production of C19-steroids. In parallel, chromaffin cells were characterized by immunohistochemical staining for chromogranin A, the major soluble protein in adrenal chromaffin granules. Large 17 alpha-hydroxylase-immunoreactive cells were found in the center of the adrenal anlagen during the 6th week of gestation. At the same developmental stage, chromaffin cells with a neuronal-like appearance occurred in the paraortic area and started to invade the adrenal primordium. Our results show that, even at week 6 gestational age, when chromaffin cells start to enter the adrenal anlagen, human adrenals already contain differentiated, 17 alpha-hydroxylase immunoreactive cortical cells which were located to the center of the primordium.

Migratory animals capable of navigating to a specific destination, and of compensating for an artificial displacement into unfamiliar territory, are thought to have a compass for maintaining their direction of travel and a map sense that enables them to know their location relative to their destination. Compasses are based on environmental cues such as the stars, the Sun, skylight polarization and magnetism, but little is known about the sensory mechanism responsible for the map sense. Here we show that the green sea-turtle (Chelonia mydas) has a map that is at least partly based on geomagnetic cues.