Aspartic Proteinase Members Secreted by the Ruminant Placenta: Specificity of Three Radioimmunoassay Systems for the Measurement of Pregnancy-associated Glycoproteins

Pregnancy‐associated glycoproteins (PAGs) isolated from the placenta of various ruminant species are enzymatically inactive members of the aspartic proteinase family. The measurement of these proteins in the maternal blood can be a good indicator of the presence of a live embryo. As certain aspartic proteinases are present in biological fluids in physiological and pathological conditions at various concentrations, it was necessary to determine the specificity of three radioimmunoassay (RIA) systems currently used for the detection of PAG molecules. Commercially available members of the aspartic proteinase family like pepsinogen, pepsin, chymosin, rennet, cathepsin D and renin were tested in a wide concentration range (10 ng/ml – 1 mg/ml). Pepsinogen cross‐reacted in RIA 1, RIA 2 and RIA 3 over 1 mg/ml, 50 μg/ml and 500 μg/ml concentrations, respectively. In the presence of pepsin, cross‐reaction was observed in RIA 1, RIA 2 and RIA 3 over 1 mg/ml, 500 μg/ml and 1 mg/ml concentrations, respectively. Chymosin and rennet could cross‐react in RIA 2 and RIA 3, while renin and cathepsin D did not decrease the binding of the tracer to antisera more, than that of the minimal detection limit. As the plasma/serum concentrations of the examined aspartic proteinases reported in the literature were outside the concentration range where cross‐reaction was observed, it can be concluded that these RIA systems were specific for the detection of PAGs in biological fluids.     

Nicotine in breast fluid of nonlactating women

Using a combination of gas chromatography, mass spectrometry, and selected ion recording techniques, we have identified nicotine and its major metabolite, continine, in the breast fluid of nonlactating women smokers. As little as 25 picograms could be measured by using the deuterated variants, [5',5'-2H]nicotine and [3,3-2H]cotinine, both as internal standards and as carriers in an inverse isotope dilution method.     

Cerumen in American Indians: genetic implications of sticky and dry types

Occurrence of sticky and dry cerumen was determined in 483 Indians from various tribes of the United States. The elevated frequencies of the allele for dry cerumen, found in Indians of pure ancestry, support the theory of the mongoloid origin of the American Indian. Potential application of cerumen quality as a marker for genetic and anthropological studies is discussed.     

Authenticity of the traditional cypriot spirit "zivania" on the basis of metal content using a combination of coupled plasma spectroscopy and statistical analysis

Sixty-eight alcoholic beverages ranging in alcoholic degree between 40 and 55 from different countries were analyzed for their 16 most abundant metal elements using inductively coupled plasma (ICP) spectroscopy. The results were analyzed statistically using two different types of analytical methods: canonical discriminant analysis and classification binary trees. The aim of this study was to investigate which of the metals analyzed constitute diagnostic parameters that establish authenticity of the traditional Cypriot spirit zivania. The two statistical methods revealed that Mg, Zn, and Cu are promising distinctive parameters capable of differentiating zivania from other spirits similar in alcoholic degree. It is believed that this differentiation in metals between the alcoholic beverages examined is related to the unique geological and climatic conditions existing on the island of Cyprus.     

Reduction in the mRNA expression of sVEGFR1 and sVEGFR2 is associated with the selection of dominant follicle in cows

The vascular endothelial growth factor (VEGF) is essential for follicular development by promoting follicular angiogenesis, as well as for the proliferation and survival of granulosa cells. The biological effects of VEGF are regulated by two membrane receptors, VEGFR1 and VEGFR2, and two soluble receptors, sVEGFR1 and sVEGFR2, which play an antagonistic role. Thus, the objective of this study was to identify the mRNA expression pattern of total VEGF, VEGFR1, VEGFR2, sVEGFR1 and sVEGFR2 in bovine preselected follicles (PRF) and post‐selected follicles (POF). The mRNA expression of these five genes in both granulosa cells (GC) and theca cells (TC) was compared between follicles classified as PRF and POF based on their diameter and on their ratio of estradiol/progesterone (E2/P4). Results showed a lower expression of mRNA of sVEGFR1 and sVEGFR2 in POF than in PRF (p < .05). Regarding the mRNA expression of total VEGF, VEGFR1 and VEGFR2, there was no difference between POF and PRF follicles (p > .05). Our results showed that the mRNA expression of VEGFR2 and sVEGFR1 was more abundant than the expression of VEGFR1 and sVEGFR2, while GC was the main source of mRNA for total VEGF. On the other hand, TC was the follicular compartment where the receptors were most expressed. Our results suggest that non‐dominant follicles maintain a greater concentration of the mRNA expression of both membrane and soluble VEGF receptors. On the other hand, follicular dominance is related to a reduction in the mRNA expression of sVEGFR1 and sVEGFR2, which may favour VEGF binding with VEGFR2 and, hence, improve the follicular health and development.     

Modeling Landscape‐scale Erosion Potential Related to Vehicle Disturbances Along the USA–Mexico Border

Decades of intensive off‐road vehicle use for border security, immigration, smuggling, recreation, and military training along the USA–Mexico border have prompted concerns about long‐term human impacts on sensitive desert ecosystems. To help managers identify areas susceptible to soil erosion from anthropogenic activities, we developed a series of erosion potential models based on factors from the Universal Soil Loss Equation (USLE). To better express the vulnerability of soils to human disturbances, we refined two factors whose categorical and spatial representations limit the application of the USLE for non‐agricultural landscapes: the C‐factor (vegetation cover) and the P‐factor (support practice/management). A soil compaction index (P‐factor) was calculated as the difference in saturated hydrologic conductivity (K_s) between disturbed and undisturbed soils, which was then scaled up to maps of vehicle disturbances digitized from aerial photography. The C‐factor was improved using a satellite‐based vegetation index, which was better correlated with estimated ground cover (r² = 0·77) than data derived from land cover (r² = 0·06). We identified 9,780 km of unauthorized off‐road tracks in the 2,800‐km² study area. Maps of these disturbances, when integrated with soil compaction data using the USLE, provided landscape‐scale information on areas vulnerable to erosion from both natural processes and human activities and are detailed enough for adaptive management and restoration planning. The models revealed erosion potential hotspots adjacent to the border and within areas managed as critical habitat for the threatened flat‐tailed horned lizard and endangered Sonoran pronghorn. Copyright © 2014 John Wiley & Sons, Ltd.     

In vitro Development of Buffalo Oocytes in Media-containing Fluids from Different Size Class Follicles

Studies were conducted to investigate the effect of supplementation of fluid from different sized class [small (SFF, < 3 mm), medium (MFF, 3-8 mm) and large (LFF, > 8 mm)] of normal and cystic (CFF) ovarian follicles in oocyte culture media on oocyte maturation rate and embryo development in vitro and to test the efficacy of follicular fluid (FF) from different size classes as a whole oocyte maturation medium. Results suggested that FF were capable of developing buffalo oocytes to embryonic stage in vitro although its efficacy was lower than that of serum. Regardless of high maturation rates after in vitro maturation (IVM) in media containing FF or IVM in whole FF, low blastocyst rates were obtained after in vitro fertilization (IVF) and culture of embryos. Follicular fluid from small follicles had significantly (p < 0.05) higher potential of developing buffalo oocytes to embryonic stage in vitro than that from medium and large follicles. Cystic FF was not capable of supporting development of buffalo oocytes in vitro.     

Effect of Semen Extender and Density Gradient Centrifugation on the Motility and Fertility of Turkey Spermatozoa

In the absence of commercially viable methods for cryopreserving turkey spermatozoa, new processing methods are required to extend the functional life of stored turkey spermatozoa for artificial insemination. The present study evaluates the efficacy of a new extender (Turkey Semen Extend) and investigates the use of density gradient centrifugation in processing turkey spermatozoa for artificial insemination. The new extender is compared with two commercially available turkey semen extenders, Beltsville Poultry Semen Extender and Ovodyl. Turkey spermatozoa in Turkey Semen Extend were still motile 20 h after collection, representing a considerable improvement over the other semen extenders (40%, 0% and 8% for Turkey Semen Extend, Beltsville Poultry Semen Extender and Ovodyl, respectively). A field trial on a commercial turkey farm showed improved fertilization rates following insemination of turkey hens with semen extended in Turkey Semen Extend (89.7%) compared with Beltsville Poultry Semen Extender (86.9%). This difference is statistically significant (p < 0.05). Processing on a density gradient, optimized for turkey spermatozoa, also increased sperm survival (50% gradient-prepared spermatozoa still motile after 18 h compared with <10% non-processed spermatozoa). Preliminary studies indicate that gradient preparation of spermatozoa may aid survival during cryopreservation.