In vitro Capacitation of Ejaculated Rabbit Spermatozoa

Prior to fertilization mammalian spermatozoa undergo physiological changes in the female reproductive tract. These changes are collectively known as capacitation. In essence capacitation is a further differentiation that the sperm cell must acquire beyond the maturational changes that it undergoes in the epididymis. Although capacitation in vitro has been easily achieved in rodents, its accomplishment in the rabbit is inconsistent and difficult. We report here successful in vitro capacitation of ejaculated rabbit sperm, assessed by in vitro and in vivo fertilization of rabbit ova. Sperm were used from pooled ejaculates collected from bucks of proven fertility since sperm collected from individual bucks resulted in significant differences in fertilization levels. Conditions favoring in vitro capacitation were: (1) extended incubation time of 12 h, (2) addition of 20% heated rabbit serum to incubation medium, and (3) an atmosphere of 5% CO2, 8% O2, and 87% N2 during incubation. In vitro capacitation of sperm under these conditions resulted in 67% fertilization compared to 89% for control sperm capacitated for 13 h in the uterus.     

Potential effects of elevated atmospheric carbon dioxide on benthic autotrophs and consumers in stream ecosystems: a test using experimental stream mesocosms

Elevated atmospheric carbon dioxide (eCO₂) has been shown to have a variety of ecosystem‐level effects in terrestrial systems, but few studies have examined how eCO₂ might affect aquatic habitats. This limits broad generalizations about the effects of a changing climate across biomes. To broaden this generalization, we used free air CO₂ enrichment to compare effects of eCO₂ (i.e., double ambient ～720 ppm) relative to ambient CO₂ (aCO₂～360 ppm) on several ecosystem properties and functions in large, outdoor, experimental mesocosms that mimicked shallow sand‐bottom prairie streams. In general, we showed that eCO₂ had strong bottom‐up effects on stream autotrophs, which moved through the food web and indirectly affected consumer trophic levels. These general effects were likely mediated by differential CO₂ limitation between the eCO₂ and aCO₂ treatments. For example, we found that eCO₂ decreased water‐column pH and increased dissolved CO₂ in the mesocosms, reducing CO₂‐limitation at times of intense primary production (PP). At these times, PP of benthic algae was about two times greater in the eCO₂ treatment than aCO₂ treatment. Elevated PP enhanced the rate of carbon assimilation relative to nutrient uptake, which reduced algae quality in the eCO₂ treatment. We predicted that reduced algae quality would negatively affect benthic invertebrates. However, density, biomass and average individual size of benthic invertebrates increased in the eCO₂ treatment relative to aCO₂ treatment. This suggested that total PP was a more important regulator of secondary production than food quality in our experiment. This study broadens generalizations about ecosystem‐level effects of a changing climate by providing some of the first evidence that the global increase in atmospheric CO₂ might affect autotrophs and consumers in small stream ecosystems throughout the southern Great Plains and Gulf Coastal slope of North America.     

Physiological Responses of Spring Durum Wheat Cultivars to Early-season Drought in a Mediterranean Environment

The Mediterranean climate of North Africa is characterized byuncertain rainfall immediately after seedling emergence, leadingto drought early in the growing season which depresses durumwheat production. However, there is limited understanding ofthe physiological basis of resistance of spring durum wheatto drought in rainfed Mediterranean regions. The objectivesof this study were to examine differences in some physiologicalcharacters among spring durum wheat cultivars in response toduration of early-season drought, and to determine the relationshipof these characters to drought resistance. In two field experiments(1995 and 1996 growing seasons) and a glasshouse experiment(1996), six spring sown durum wheat cultivars were evaluatedunder four water regimes: well irrigated and three differentwater deficits from emergence until the onset of tillering,mid-tillering or at the end of tillering. Cultivars differedin their response. Decreases in photosynthesis soon after droughtstress was imposed resulted mainly from reduced stomatal conductance.Continued water deficits also reduced mesophyll photosyntheticactivity. Possible factors determining the drought-resistanceof a cultivar are lower sensitivity of CO₂exchange rate, netCO₂uptake to water loss ratio, stomatal resistance, relativewater content and greater osmotic adjustment under stress. Furthermore,there is sufficient intraspecific variation in these physiologicalattributes to suggest their use as selection tools.Copyright1998 Annals of Botany Company Wheat;Triticum durumDesf.; early-season drought; physiological responses.     

DNA phylogeny supports revised classification of Salmothymus obtusirostris

Salmothymus obtusirostris (soft-muzzled trout) is endemic to the South Adriatic drainage. Owing to its unusual appearance, which resembles both trout and grayling, it has been initially classified as a separate genus. However, this classification is ambiguous and has never been firmly established. We have studied mtDNA (control region and cytochrome b gene) and nuclear DNA (a part of LDH C*1 gene) variation between soft-muzzled trout from the upper part of the River Neretva, Bosnia and Herzegovina, and other salmonid representatives in order to examine how the current classification is congruent with molecular data. On the basis of sequence identity of mtDNA control region among several genera (i.e. Salmo , Oncorhynchus , Salvelinus , Acantholingua , Brachymystax , Thymallus and Coregonus ) a close relationship between Salmothymus , Salmo and Acantholingua was established. Phylogenetic analysis on a combined data set of mitochondrial and nuclear DNA, supported by 100% bootstraping, indicated that S. obtusirostris and A. ohridana are sister taxa which exhibit a closer relationship to S. trutta than to S. salar . This finding refutes the current classification, which recognizes S. obtusirostris as separate genus, and instead suggests its reclassification on the species level as Salmo obtusirostris .  © 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 2002, 77 , 399–411.     

Embryonic Appearance of α, β, and γ Crystallins in the Periodic Albinism (ap) Mutant of Xenopus laevis

The appearance of the crystallins during lens development in the periodic albinism (a^p/a^p) mutant of Xenopus laevis has been studied. Using antibodies specific for total crystallins, α+β crystallins, and γ crystallins in the immunofluorescence technique, the first positive reaction for all could be demonstrated in the Nieuwkoop-Faber Stage 31 lens rudiment. The antibody to α+β crystallins exhibited differences in intensity from cell to cell in the early rudiment, while the reaction to the other antibodies was uniform throughout the rudiment. As lens differentiation progressed, immunofluorescence was restricted in all cases to the lens fiber area, up to and including Nieuwkoop-Faber Stage 45. The lens epithelium of the one-year-old adult a^p/a^p was positive, however, for total lens crystallins.
These results are at variance with earlier studies on lens development and the crystallins in wild-type (+/+) X. laevis , where a positive reaction for y and total crystallins could be detected earlier, and in the lens epithelium of Nieuwkoop-Faber Stage 41 embryos for total lens crystallins. That this divergence in the mutant is due to a pleiotropic effect or directly to the inductive failure of the endomesoderm to initiate melanogenesis, is discussed.     

Doping in sports and its spread to at-risk populations: an international review

Doping is now a global problem that follows international sporting events worldwide. International sports federations, led by the International Olympic Committee, have for the past half century attempted to stop the spread of this problem, with little effect. It was expected that, with educational programs, testing, and supportive medical treatment, this substance-abusing behavior would decrease. Unfortunately, this has not been the case. In fact, new, more powerful and undetectable doping techniques and substances are now abused by professional athletes, while sophisticated networks of distribution have developed. Professional athletes are often the role models of adolescent and young adult populations, who often mimic their behaviors, including the abuse of drugs. This review of doping within international sports is to inform the international psychiatric community and addiction treatment professionals of the historical basis of doping in sport and its spread to vulnerable athletic and non-athletic populations.