An overview of the serpin superfamily

Serpins are a broadly distributed family of protease inhibitors that use a conformational change to inhibit target enzymes. They are central in controlling many important proteolytic cascades, including the mammalian coagulation pathways. Serpins are conformationally labile and many of the disease-linked mutations of serpins result in misfolding or in pathogenic, inactive polymers.     

Studies of the pathogenesis of bovine pestivirus-induced ovarian dysfunction in superovulated dairy cattle

Two experiments (Experiment I, n=12 Holstein-Friesian heifers; Experiment II, n=8 Jersey cows) were conducted to investigate the pathogenesis of bovine pestivirus-induced ovarian dysfunction in cattle. In both experiments the cattle were superovulated with twice daily injections of a porcine pituitary extract preparation of follicle stimulating hormone (FSH-P), for 4 days commencing on Day 10+/-2 after a presynchronised oestrus. The heifers received a total dose of 30 mg and the cows 32 mg of FSH-P. Prostaglandin F(2alpha) (PGF(2alpha)) was administered 48 h after commencement of superovulation and all cattle were artificially inseminated (AI) between 48 and 66h after PGF(2alpha) treatment. In both experiments bovine pestivirus seronegative cattle (Experiment I, n=6; Experiment II, n=4) were inoculated intranasally with an Australian strain of non-cytopathogenic bovine pestivirus (bovine viral diarrhoea virus Type 1) 9 days prior to AI. Bovine pestivirus infection was confirmed by seroconversion and/or virus isolation in all of the inoculated cattle, consistent with a viremia occurring approximately between Day 5 prior to AI and the day of AI. Ovarian function was monitored in both experiments by daily transrectal ultrasonography and strategic blood sampling to determine progesterone, oestradiol-17beta, luteinising hormone (LH) and cortisol profiles. Non-surgical ova/embryo recovery was performed on Day 7 after AI. In Experiment II half the cattle were slaughtered on Day 2 and the remainder on Day 8 after AI, and the ovaries submitted for gross and histopathological examination including immunohistochemistry to demonstrate the presence of bovine pestivirus antigen. In both studies, comparisons were made between infected and confirmed uninfected (control) animals. Overall the bovine pestivirus infected cattle had significantly lower (P<0.05) ova/embryo recovery rates compared to the control cattle. There was evidence of either an absence (partial or complete) of a preovulatory LH surge or delay in timing of the LH peak in the majority (90%) of infected heifers and cows, and histologically, there was evidence of non-suppurative oophoritis with necrosis of granulosa cells and the oocyte in follicles from the infected cows. By contrast only 20% of the control heifers and cows had evidence of absence of a pre-ovulatory LH surge. These experiments collectively demonstrate that bovine pestivirus infection during the period of final growth of preovulatory follicles may result in varying degrees of necrosis of the granulosa cells with subsequent negative effects on oestradiol-17beta secretion which in turn negatively affects the magnitude and/or timing of the preovulatory LH surge.     

Microbial mats on the Orkney Islands revisited: microenvironment and microbial community composition

The microenvironment and community composition of microbial mats developing on beaches in Scapa Flow (Orkney Islands) were investigated. Analysis of characteristic biomarkers (major fatty acids, hydrocarbons, alcohols, and alkenones) revealed the presence of different groups of bacteria and microalgae in mats from Waulkmill and Swanbister beach, including diatoms, Haptophyceae, cyanobacteria, and sulfate-reducing bacteria. These analyses also indicated the presence of methanogens, especially in Swanbister beach mats, and therefore a possible role of methanogenesis for the carbon cycle of these sediments. High amounts of algal lipids and slightly higher numbers (genera, abundances) of cyanobacteria were found in Waulkmill Bay mats. However, overall only a few genera and low numbers of unicellular and filamentous cyanobacteria were present in mats from Waulkmill and Swanbister beach, as deduced from CLSM (confocal laser scanning microscopy) analysis. Spectral scalar irradiance measurements with fiber-optic microprobes indicated a pronounced heterogeneity concerning zonation and density of mainly anoxygenic phototrophs in Swanbister Bay mats. By microsensor and T-RFLP (terminal restriction fragment length polymorphism) analysis in Swanbister beach mats, the depth distribution of different populations of purple and sulfate-reducing bacteria could be related to the microenvironmental conditions. Oxygen, but also sulfide and other (inorganic and organic) sulfur compounds, seems to play an important role in the stratification and diversity of these two major bacterial groups involved in sulfur cycling in Swanbister beach mats.     

Light-induced carotenogenesis in Myxococcus xanthus: DNA sequence analysis of the carR region

The carR region encodes a light-inducible promoter, a negative regulator of the promoter and a trans-acting activator that controls the light-inducible Myxococcus xanthus carotenoid biosynthesis regulon. DNA sequence analysis revealed, downstream of the promoter, three translationally coupled genes, carQ, carR and carS. Sequencing of mutations demonstrated that carR encoded the negative regulator and was an integral membrane protein. Mutant construction and sequencing revealed that carS was the trans-acting activator and that carQ was a positive regulator of the promoter. Neither gene encodes proteins with known sequence-specific DNA-binding motifs. The sequence of the light-inducible promoter region, identified by primer extension analysis, showed similarity to the consensus sequence of the Escherichia coli stress response (‘heat-shock’) promoters.     

A field investigation of the effects of bovine viral diarrhea virus infection around the time of insemination on the reproductive performance of cattle

During a study of methods of synchronizing estrus in Bos indicus cattle, blood was collected from 169 heifers and 38 cows 2 to 3 days prior to artificial insemination (AI), and then again at Day 51 and Day 210 after AI to determine the incidence of infection with bovine viral diarrhea (BVD) virus. Prior to insemination 53 and 68% of the cows and heifers, respectively, were seronegative to the BVD virus. At Day 51 after AI, 70 and 32% of the seronegative cows and heifers, respectively, had seroconverted; but between Day 51 and Day 210, only 17 and 3% of the seronegative cows and heifers, respectively, had seroconverted. The Day- 51 pregnancy rate of cows which were immune (seropositive) to BVD virus infection at the time of AI was similar to the rate of the cows which became infected around the time of AI. However, the pregnancy rate of the immune heifers (44%, n=54) was significantly (P=0.04) greater than the rate of the heifers which became infected around the time of AI (24%, n=37). It was concluded that infection of susceptible females with BVD virus around the time of AI may significantly lower the pregnancy rate.     

Effect of exogenous progestogens on plasma concentrations of the oxytocin-associated neurophysin and 13,14-dihydro-15-keto-prostaglandin F in intact and ovariectomized ewes over the time of expected luteal regression

Plasma concentrations of the prostaglandin F metabolite 13,14-dihydro-15-keto-prostaglandin F (PGFM), the oxytocin-associated neurophysin (OT-N) and progesterone were monitored by radioimmunoassay (RIA) in five ewes sampled from the jugular vein at hourly intervals between 0700-1900 h from Days 12-16 of the estrous cycle. These hormones were also determined in plasma samples collected at similar times from five intact and five ovariectomized ewes given twice daily injections of medroxyprogesterone acetate (MPA) over Days 10 to 20 after the last observed estrus. In both the control and intact MPA-treated ewes, coincident surges of OT-N and PGFM were observed in jugular plasma during the time of luteal regression. No significant differences were noted in the number and amplitude of OT-N and number of PGFM peaks between the control and intact MPA-treated animals, although in the latter the amplitude of the PGFM peaks was significantly reduced (P less than 0.01). No marked surges in the plasma concentrations of PGFM or OT-N were observed in the ovariectomized ewes given exogenous MPA. This latter finding is consistent with previous proposals, suggesting that the ovaries are a major source of oxytocin in the ewe. In addition, the observation that exogenous progestogens in the intact ewes did not influence the number and peak height of the OT-N surges, indicates that a fall in progesterone levels during the normal cycle is not obligatory for oxytocin release although it may facilitate the release of uterine PGF2 alpha.     

Total protein content and protein synthesis within pre-elongation stage bovine embryos

Protein content was measured in zona-free bovine oocytes and pre-elongation stage embryos, following in vitro maturation, fertilisation, and then culture in Synthetic Oviduct Fluid medium supplemented with amino acids and 8 mg ml^-1 bovine serum albumin (BSA). Values (ng embryo^-1) of 122 ± 7.8, 137 ± 8.6, 111 ± 8.8, 115 ± 10.4, 139 ± 9.0 and 152 ± 10.1 were obtained for zona-free mature oocytes, 2-cell (day 2), 8-cell (day 3), compact morula (day 6), blastocyst (day 7), and expanded blastocyst (day 8) stage embryos, respectively. The protein content of day 7 zona-enclosed blastocysts was 337 ± 58.0 ng embryo^-1. These values suggest that prior to compaction and blastulation, the early cleavage stage bovine embryo has a higher rate of protein degradation than that of synthesis. Net growth is observed only after initiation of compaction. The protein content of day 7 blastocysts was measured in embryos following in vitro production and culture in the same media supplemented with either 0.5% w/v polyvinyl alcohol (PVA), 8 mg ml^-1 BSA, 8 mg ml^-1 BSA and further supplemented with 10% fetal calf serum (FCS) from the beginning of culture (FCS-D1), 8 mg ml^-1 BSA and 10% FCS from the fourth day of culture (day 5 of development) or from in vivo-derived day 7 blastocysts. Protein content was significantly (P< 0.05) lower in PVA-cultured embryos than other treatments. To determine if this difference in PVA-cultured embryos was due to a difference in the rate of protein synthesis, comparisons were made between day 7 embryos derived from BSA-culture and either PVA-culture, FCS-D1 culture or in vivo-derived embryos. Despite differences in diameter, no significant difference was observed in the incorporation of L-[2,3,4,5,6-³H]-phenylalanine into the TCA-precipitable fraction in any of the three comparisons made. However, incubation in the presence of FITC-labelled BSA or β-casein and examination under either fluorescence or confocal microscopy revealed that protein in the extra-embryonic environment was actively taken up by the trophectoderm of day 7 blastocysts, most likely by endocytosis. These results suggest that exogenous protein is an important nutritive source, probably maintaining intracellular amino acid pools. Results obtained from the production of embryos in protein-free medium should be viewed with the knowledge that such embryos differ metabolically from those embryos grown in the presence of protein, including in vivo-derived embryos. Mol. Reprod. Dev. 50:139–145, 1998. © 1998 Wiley-Liss, Inc.     

Cyclicity in the fossil record mirrors rock outcrop area

In a recent article, Rohde & Muller (Rohde & Muller 2005 Nature 434, 208-210) identified a strong 62 Myr cyclicity in the history of marine diversity through the Phanerozoic. The data they presented were highly convincing, yet they were unable to explain what process might have generated this pattern. A significant correlation between observed genus-level diversity (after removal of long-term trends) and the amount of marine sedimentary rock measured at a surface outcrop in Western Europe is demonstrated. This suggests that cyclicity originates from long-term changes in sedimentary depositional and erosional regimes, and raises the strong possibility that the cyclicity apparent in the record of marine fossils is not a biological signal but a sampling signal.