Methyl‐β‐Cyclodextrin Improves Sperm Capacitation Status Assessed by Flow Cytometry Analysis and Zona Pellucida‐Binding Ability of Frozen/Thawed Bovine Spermatozoa

Mammalian sperm undergo a series of biochemical transformations in the female reproductive tract that are collectively known as capacitation. Cyclodextrins added to the sperm culture medium have been described to induce in vitro sperm capacitation, enabling its use in protein‐free media. However, the additive capacitating effect of methyl‐β‐cyclodextrin (MβCD) in the medium containing bovine serum albumin (BSA) is unknown in the bovine species. In this study, we evaluated the effects of incubating frozen–thawed bovine spermatozoa in a BSA‐containing medium supplemented with MβCD on different sperm quality and functional parameters. Sperm viability decreased with the addition of MβCD in a dose‐dependent manner (p < 0.05), and DNA damage could be observed but only with the highest concentration of MβCD. However, pre‐incubation of spermatozoa in MβCD‐supplemented medium improved the capacitation status as assessed by the increase in plasma membrane fluidity, intracellular calcium concentration, induced acrosome reactivity and zona pellucida (ZP)‐binding ability (p < 0.05). Thus, we conclude that MβCD supplementation is able to enhance the capacitation status of frozen–thawed bovine spermatozoa cultured in capacitation medium containing BSA and could result in a valid strategy for its application on artificial reproductive technologies such as in vitro fertilization or intracytoplasmic sperm injection.     

Magnetic field effect on picosecond electron transfer

The recombination dynamics of a transition metal redox system monitored by femtosecond pump-probe spectroscopy are shown to be sensitive to high magnetic fields at times shorter than 10 picoseconds. The effect, based on coherent population beats of different spin states, is quantitatively accounted for and allows direct access to spin relaxation rates far beyond the time resolution of the fastest electron paramagnetic resonance technique. The presence of this ultrafast magnetic field effect helps in understanding complex reaction schemes in transition metal chemistry, which occur in a wide range of fields, such as bioinorganic chemistry and catalysis.     

Biochemical and Protein Profile of Alpaca (Vicugna pacos) Uterine Horn Fluid During Early Pregnancy

South American camelids show high embryo loss rate, during the first 60 days of pregnancy. One of the factors which may be related to this situation is that over 98% of the embryos implant in the left uterine horn (LUH) even though both ovaries contribute similarly to ovulation. There is scarce information about the uterine environment of female camelids at any physiological state that could explain the capability of the LUH to attract the embryo and maintain pregnancy. We describe, for the first time, the biochemical and protein profile of uterine fluid (UF), addressing the right and LUH environment in non‐pregnant and pregnant alpacas. Different substrates, electrolytes and metabolites were assayed in both uterine horn fluids. Small changes were observed in glucose and total protein levels, which were more noticeable during pregnancy. In addition, 10 specific proteins were found in the left horn fluid in 5‐week‐pregnant alpacas, and two protein bands were identified in non‐pregnant alpaca right horn fluid. These results would provide basic information for identification of possible markers for pregnancy diagnosis, reproductive diseases and hormone‐treated animals evaluation and hence contributing to improve the pregnancy rate.     

Subcritical water extraction of antioxidant compounds from rosemary plants

Subcritical water extraction at several temperatures ranging from 25 to 200 degrees C has been studied to selectively extract antioxidant compounds from rosemary leaves. An exhaustive characterization of the fractions obtained using subcritical water at different temperatures has been carried out by LC-MS, and the antioxidant activities of the extracts have been measured by a free radical method (DPPH). Results indicate high selectivity of the subcritical water toward the most active compounds of rosemary such as carnosol, rosmanol, carnosic acid, methyl carnosate, and some flavonoids such as cirsimaritin and genkwanin. The antioxidant activity of the fractions obtained by extraction at different water temperatures was very high, with values around 11.3 microg/mL, comparable to those achieved by SFE of rosemary leaves. A study of the effect of the temperature on the extraction efficiency of the most typical rosemary antioxidant compounds has been performed.     

Trees and farming in the dry zone of southern Honduras II: the potential for tree diversity conservation

The potential of the dry zone agro-ecosystem of southern Honduras to contribute to the conservation of Mesoamerican dry forest tree diversity is evaluated. Four rural communities containing eight land uses were surveyed using rapid botanical sampling resulting in the identification of 241 tree and shrub species. As a result of ordination analysis, it is concluded that the land uses are relatively similar in their species composition, particularly maize fields (milpas), fallows, pastures and woodlots, because of the predominance of natural regeneration. Therefore all land uses might contribute to local tree diversity conservation. Those land uses in which planting also contributes to diversity, home gardens (solares) and orchards, are more distinct; however the tree species found there are widespread and often exotics and thus not the usual focus of conservation measures. Across the landscape the total complement of species considered a global priority for biodiversity conservation is very low and therefore this agro-ecosystem does not represent a good place in which to implement dry forest tree diversity conservation programmes. Instead its value is likely to be in the contribution that tree diversity makes to rural livelihoods. Particular consideration is given to Swietenia humilis Zucc. (small leaved mahogany) and its status as a threatened species is questioned because of its abundance within this landscape and its wide distribution. This revised version was published online in June 2006 with corrections to the Cover Date.     

Fibrocartilaginous embolic myelopathy in miniature schnauzers.

In large- and giant-breed dogs, fibrocartilaginous embolic myelopathy (FCEM) is a well-recognized syndrome of acute spinal cord infarction caused by embolization of fibrocartilage. The miniature schnauzer is reportedly the most frequently affected small breed, although clinical data from only six miniature schnauzers with FCEM is available in the literature. The purposes of this study were to determine the relative frequency of FCEM compared to other causes of myelopathy in miniature schnauzers, to characterize the clinicopathological features of FCEM in 38 miniature schnauzers, and to directly compare FCEM and intervertebral disk herniation in miniature schnauzers with respect to age at diagnosis; gender; neuroanatomical localization; and progression, asymmetry, and severity of neurological deficits. Fibrocartilaginous embolic myelopathy was the most common cause of myelopathy in miniature schnauzers. Age at diagnosis, asymmetry and severity of neurological deficits, and lack of progression of clinical signs after 24 hours assisted in distinguishing FCEM from intervertebral disk herniation. Fibrocartilaginous embolic myelopathy-related mortality in miniature schnauzers was significantly lower than mortality rates reported for affected large and giant breeds. Only 22% of miniature schnauzers were euthanized because of their disease, although the vast majority of survivors failed to achieve complete neurological recovery.     

Intermediate Filament Proteins Expression and Carbohydrate Moieties in Trophoblast and Decidual Cells of Mature Cat Placenta

The aim of this study was to characterize cytoskeletal intermediate filament proteins and glycoconjugates of syncytiotrophoblast, cytotrophoblast and decidual cells of feline endotheliochorial placenta. Samples from 12 normal pregnant female cats, after 45 ± 5 days of gestation, were obtained removing the uterine horns by hysterectomy. Sections were processed for routine observation and for immunohistochemistry using anticytokeratin, antivimentin and antidesmin antibodies. In addition, lectin histochemistry was performed using a panel of several biotinylated lectins to characterize glycosides expression profile. Cytotrophoblast and syncytiotrophoblast showed immunoreactivity only with acidic and basic cytokeratins. Decidual cells were only positive to vimentin, consistent with their origin from endometrial fibroblasts. Trophoblast expressed a broad population of glycans, highly exposing terminal N‐acetyl glucosamine residues and non‐sialylated galactose and N‐acetyl galactosamine oligomers. Oligosaccharides bound by Phaseolus vulgaris erythroagglutinin were the only highly branched N‐linked residues evidenced in cats, and they were restricted to the syncytium. Unlike results reported on humans, mice and rats on lectin affinity of decidual cells, sialid acids and complex N‐linked oligosaccharides were not demonstrated in cats. Glycosylation of proteins determines many of their final properties, thus becoming essential for the embryo‐maternal dialogue during implantation and placentation. Changes in glycosylation pattern have been related to pathological pregnancies in other species. Hence, the knowledge about glycosylation profile of the normal cat placenta may lead to a better understanding of both normal and pathological reproductive events.