Conservation Genetics of the East Pacific Green Turtle (Chelonia mydas) in Michoacan, Mexico

The main continental nesting rookeries of the east Pacific green turtle (EPGT), Chelonia mydas, on the Michoacan (Mexico) coast suffered drastic population declines following intense exploitation in the 1960s--1970s with annual abundance of nesting females plummeting from about 25,000 to an average of about 1400 between 1982 and 2001. Analyses of data from three nDNA microsatellite loci and 400 bp mtDNA control region sequences from a total of 123 nesting females sampled from four Michoacan rookeries found no evidence of population sub-structuring. The recent order of magnitude reduction in the population size shows no apparent impact on genetic diversity in either control region sequences (overall h = 0.48; pi = 0.0036) or microsatellite loci (overall Na = 20.8; Hexp = 0.895). Our estimates of annual effective female population size (Nef; from theta = 2Nemicron) of 1.9-2.3 x 10(3), in spite of being an order of magnitude below historical records, appear to be sufficient to allow recovery of this population without significant loss of genetic diversity. These findings highlight the importance of continued conservation to reverse the decline of this population before it becomes vulnerable to genetic erosion.     

Inhibitory effects on phospholipase A2 and antivenin activity of melanin extracted from Thea sinensis Linn

Antivenin activity of melanin extracted from black tea (MEBT) was reported for the first time. The antagonistic effect of MEBT was evaluated for Agkistrodon contortrix laticinctus (broadbanded copperhead), Agkistrodon halys blomhoffii (Japanese mamushi), and Crotalus atrox (western diamondback rattlesnake) snake venoms administered i.p. to ICR mice. MEBT was injected i.p. immediately after the venom administration in dose of 3 mg per mouse in the same place of venom injection. MEBT demonstrated neutralization effect against all venoms tested. The greatest antivenin effect of MEBT was found against Japanese mamushi snake venom. In this case, half the mice died within 2.5 +/- 0.7 h after injection of 0.9 mg/kg of venom. An immediate injection of MEBT substantially reduced the toxic effect of venom and extended time at the 50% level of survival up to 52.3 +/- 2.3 h. The antivenin activity of MEBT is due to chelating of Ca++ and non-specific binding of phospholipase A2. The inhibitory effect of MEBT on phospholipase A2 assessed for different venoms was similar to that obtained with pure enzyme. Low toxicity of MEBT in combination with its antagonistic activity against different venoms may allow effective life-saving treatment against snakebites. Such application of MEBT is important when identification of the snake is impossible or if specific treatment is unavailable.     

酸提高绿茶提取物对脂肪酸合酶的抑制活性

脂肪酸合酶和肥胖、癌症等人类重大疾病相关.获取高活性脂肪酸合酶抑制剂有重要应用价值.50％乙醇的绿茶提取物具有抑制脂肪酸合酶和经口服降低大、小鼠体重的功能.该提取物在酸的作用下可明显地提高其抑制脂肪酸合酶的能力.采用1 mol/L硫酸或盐酸在100℃下处理绿茶提取物,发现该提取物对脂肪酸合酶的抑制活性随时间逐步提高.在25℃至120℃温度下,提高处理温度使抑制活性提高的速度加快和程度加大. 用1 mol/L盐酸120℃处理35 min或3 mol/L盐酸100℃处理30 min可使绿茶提取物抑制脂肪酸合酶的半抑制浓度下降20倍左右,达到1 μg/ml以下.用pKa值4.76至1.27的1 mol/L浓度的4种有机酸在100℃作用150 min使绿茶提取物抑制活性分别提高到1.48至5.84倍.提高酸的浓度也使被作用的提取物的抑制活性提高的更多.表现抑制活性提高的速度及程度和氢离子浓度正相关.判断绿茶提取物在酸作用下抑制活性的提高来源于其中所含儿茶素在氢离子作用下发生了化学变化.初步实验表明其变化过程比较复杂,确定其分子机制还需进一步的研究工作.     

Metabolic profiling in validation of plasma biomarkers for green tea polyphenols

Green tea polyphenols (GTP) effectively protect against chronic diseases in various animal models but human studies have been inconclusive. GTP components and metabolites in body fluids have been suggested as potential biomarkers, but validation of these biomarkers has rarely been done in human populations. A randomized, double-blinded, and placebo-controlled phase IIa chemoprevention study with GTP was conducted in 120 human subjects for 3 months. To validate GTP biomarker profiles, plasma samples were collected at baseline, 1-month, and 3-month and were analyzed by HPLC-Coularray electrochemical detection (ECD) for specific GTP components as well as for non-targeted metabolites. The levels of 2 GTP components, epigallocatechin-3-gallate (EGCG) and epicatechin-3-gallate (ECG), were homogenous at baseline (p > 0.45) but were significantly elevated (p < 0.01) by GTP treatment. Metabolic profiling identified 106 metabolites, and 56 of them were chosen to construct discriminant functions (DFs) based on the data at 3 months. The DFs clearly separated the placebo, 500 mg GTP, and 1000 mg GTP groups with an accuracy rate of 97.3%. When the DFs were applied to the combined baseline and 1-month data, the accuracy rate was 62.9% in classifying subjects into the 3 intervention groups. DFs derived from 1-month data showed similar results. Overall, this study validated plasma EGCG and ECG as reliable biomarkers for GTP consumption, and found metabolic profiles effective in discriminating different GTP dosages.     

Development of six microsatellite loci for black mangrove (Avicennia germinans)

We developed six new microsatellite loci for the black mangrove (Avicennia germinans), an important member of wetland communities worldwide. Loci showed moderate to high polymorphism and a survey of four locations [Puerto Rico (Jobos Bay and Luquillo), Mexico, French Guyana] revealed clear regional (and local) population structure. All populations were genetically distinct and the two continental populations showed much higher diversity than the two insular Puerto Rican locations. These loci complement those recently published by Nettel et al. (2005 ) and promise to be valuable for characterizing local and regional population dynamics in the black mangrove.     

Impact of green tea on oxidative stress induced by ammonium metavanadate exposure in male rats

Transitional metals, as vanadium, are known to exert noxious effects by generating oxidative stress. Addition of antioxidants in the diet could decrease the cytotoxic effect related to the oxidative stress. The present study, carried out in Wistar rats, is a contribution to the evaluation of protective effects of green tea Camellia sinensis, which is known to be rich in antioxidant compounds (polyphenols...). Rats were divided into four groups: (C) was control, (V) was given ammonium metavanadate (AMV), (TH) was given herbal tea as drink (66 g/l) and TH + V was given tea and metavanadate. Group (TH) was given herbal tea one month before vanadium treatment. Metavanadate was daily i.p. injected (5 mg NH4VO3/kg body weight) for 10 days. (C) and (TH) groups received i.p. injections of 0.9% NaCl during the same period. Changes in lipid peroxidation levels (TBARS) in kidney, liver and testes, serum concentrations of vitamins E and A and superoxidismutase (SOD) and catalase (CAT) activities in blood cells were determined. One month pre-treatment with green tea, followed by 10 days of treatment (TH) did not change TBARS in liver and testes as compared to controls, but induced a clear decrease of TBARS in kidneys. Intraperitoneal administration of AMV to rats (V) induced a time-dependant increase of TBARS in kidney, liver and testes that was lowered in rats (V + TH) drinking tea. Vitamin E concentrations were found to be drastically decreased from day 1 to 10 in rats (V). Vitamin A concentration was decreased at day 10 only. Drinking tea lowered AMV inhibitory effects in rats (V + TH), and conversely an increase of vitamins A and E concentrations were found at day 10. SOD and catalase activities were found increased in the blood cells from day 1 to day 5 and conversely decreased at day 10. In contrast, associated to green tea, AMV did not affect SOD and catalase activities compared to controls.     

Influence of weather conditions on the flight of migrating black storks

This study tested the potential influence of meteorological parameters (temperature, humidity, wind direction, thermal convection) on different migration characteristics (namely flight speed, altitude and direction and daily distance) in 16 black storks (Ciconia nigra). The birds were tracked by satellite during their entire autumnal and spring migration, from 1998 to 2006. Our data reveal that during their 27-day-long migration between Europe and Africa (mean distance of 4100 km), the periods of maximum flight activity corresponded to periods of maximum thermal energy, underlining the importance of atmospheric thermal convection in the migratory flight of the black stork. In some cases, tailwind was recorded at the same altitude and position as the birds, and was associated with a significant rise in flight speed, but wind often produced a side azimuth along the birds'' migratory route. Whatever the season, the distance travelled daily was on average shorter in Europe than in Africa, with values of 200 and 270 km d⁻¹, respectively. The fastest instantaneous flight speeds of up to 112 km h⁻¹ were also observed above Africa. This observation confirms the hypothesis of thermal-dependant flight behaviour, and also reveals differences in flight costs between Europe and Africa. Furthermore, differences in food availability, a crucial factor for black storks during their flight between Europe and Africa, may also contribute to the above-mentioned shift in daily flight speeds.     

防御活性氧保护系统在黑豆抗旱生理中的响应

以黑豆幼苗为材料,研究了水分胁迫下其非酶促保护物质和保护酶系对活性氧的清除作用。结果显示:黑豆皮红色素(17.36%)对DPPH.的清除率高达93.6%,可溶性糖(7.227%)对O2-与.OH的清除率分别为47.71%与68.16%,谷胱甘肽(0.8033mg/g)对O2-与.OH有较明显的清除作用,三种非酶促保护物质随着浓度的升高其清除能力呈逐渐增强的趋势;在水分胁迫下,黑豆幼苗超氧化物歧化酶(33~14 U.mg-1.min-1.2~6d-1),过氧化氢酶(2.87~2.01 mg.mg-1.min-1.3~6d-1)与过氧化物酶(31~14 mg.mg-1.min-1.3~6d-1)三种保护酶活性呈现先升高后降低的变化趋势。研究表明:黑豆体内清除活性氧防御系统作用十分明显,且具有较强的抗氧化、抗衰老、抗逆能力。     

黑豆皮中红色色素萃取工艺的响应面优化

利用逐因子试验和Box-Behnken试验对黑豆皮中红色色素萃取工艺进行了优化研究,获得了工艺优化的数学模型y=3.85+0.43x1+0.05x2-0.32x3-0.12x1x2+0.063x1x3+1.0E-0.02x2x3-0.58x12-0.49x22-0.34x32,分别对自变量求偏导得到优化工艺条件:x1=0.346,x2=0.006988,x3=-0.44,通过转换得到实际工艺条件,即萃取温度X1=83.46℃、萃取时间X2=30.07 m in、料液比X3=1:35.6(w/v)。在最优化条件下模型预测OD512为3.997,验证试验结果为3.85±0.01(n=3),试验证明,逐因子试验和Box-Behnken试验联用可以很好的应用于黑豆皮红色色素的萃取工艺中。     

Changing sources of soil respiration with time since fire in a boreal forest

Radiocarbon signatures (Δ¹⁴C) of carbon dioxide (CO₂) provide a measure of the age of C being decomposed by microbes or respired by living plants. Over a 2‐year period, we measured Δ¹⁴C of soil respiration and soil CO₂ in boreal forest sites in Canada, which varied primarily in the amount of time since the last stand‐replacing fire. Comparing bulk respiration Δ¹⁴C with Δ¹⁴C of CO₂ evolved in incubations of heterotrophic (decomposing organic horizons) and autotrophic (root and moss) components allowed us to estimate the relative contributions of O horizon decomposition vs. plant sources. Although soil respiration fluxes did not vary greatly, differences in Δ¹⁴C of respired CO₂ indicated marked variation in respiration sources in space and time. The ¹⁴C signature of respired CO₂ respired from O horizon decomposition depended on the age of C substrates. These varied with time since fire, but consistently had Δ¹⁴C greater (averaging ～120‰) than autotrophic respiration. The Δ¹⁴C of autotrophically respired CO₂ in young stands equaled those expected for recent photosynthetic products (70‰ in 2003, 64‰ in 2004). CO₂ respired by black spruce roots in stands >40 years old had Δ¹⁴C up to 30‰ higher than recent photosynthates, indicating a significant contribution of C stored at least several years in plants. Decomposition of O horizon organic matter made up 20% or less of soil respiration in the younger (<40 years since fire) stands, increasing to ～50% in mature stands. This is a minimum for total heterotrophic contribution, since mineral soil CO₂ had Δ¹⁴C close to or less than those we have assigned to autotrophic respiration. Decomposition of old organic matter in mineral soils clearly contributed to soil respiration in younger stands in 2003, a very dry year, when Δ¹⁴C of soil respiration in younger successional stands dropped below those of the atmospheric CO₂.