Post-cervical compared with cervical insemination in gilts: Reproductive variable assessments.

The aim with this study was to compare cervical (CAI; 3 × 109 spermatozoa/90 mL) and post-cervical (PCAI; 1.5 × 109 spermatozoa/45 mL) artificial insemination (AI) techniques for frequency of incidences (unsuccessful or difficult probe passage, backflow, metritis and bleeding), values for reproductive variables and duration of the procedure in gilts. There were 644 gilts (255-270 days old, weighing 150 ±â€¯5 kg) randomly assigned to PCAI (n = 320) and CAI (n = 324) groups. In total, there were 957 and 958 artificial inseminations performed in the CAI and PCAI groups, respectively (2-4 AIs/gilt). The frequency of unsuccessful or difficult PCAI probe passage/AI was 14.6% (140/958), therefore, there was a 85.7% probe passage success/AI rate (818/958). The semen backflow frequency/AI was less with PCAI than CAI (4.3% compared with 8.2%, P <  0.001). With the PCAI group, there were only a few cases of bleeding (11/958: 1.1% /AI) with no difference between the CAI and PCAI groups (P = 0.224). In gilts (n = 72) where there was not passage of the PCAI probe (72/320; 22.5%) there was use of CAI, (M, mixed group). For the CAI, PCAI and M groups, there were similar values for positive pregnancy diagnosis, farrowing rates and prolificacy (P > 0.05). The average duration for AI was shorter in the PCAI (2.34 ± 0.809 min) than CAI (4.77 ± 1.059 min) group, and it was longer in the M group (7.48 ± 2.454 min; P < 0.050). The PCAI procedure, therefore, is recommended for AI of gilts.

Biomolecular interaction analysis in functional proteomics.

To understand the function of highly complex eukaryotic tissues like the human brain, in depth knowledge about cellular protein networks is required. Biomolecular interaction analysis (BIA), as a part of functional proteomics, aims to quantify interaction patterns within a protein network in detail. We used the cAMP dependent protein kinase (PKA) as a model system for the binding analysis between small natural ligands, cAMP and cAMP analogues, with their physiological interaction partner, the regulatory subunit of PKA. BIA comprises a variety of methods based on physics, biochemistry and molecular biology. Here we compared side by side real time SPR (surface plasmon resonance, Biacore), a bead based assay (AlphaScreen), a fluorescence based method (Fluorescence polarisation) and ITC (isothermal titration calorimetry). These in vitro methods were complemented by an in cell reporter assay, BRET(2) (bioluminescence resonance energy transfer), allowing to test the effects of cAMP analogues in living cells.

Environmental pesticide exposure in Honduras following hurricane Mitch.

OBJECTIVE: To investigate whether environmental contamination occurred in the wake of hurricane Mitch (30-31 October 1998), we conducted a population-based cross-sectional household survey in the barrio of Istoca, Department of Choluteca, Honduras. The goals were to evaluate chemical contamination of potable water and the extent of human exposure to chemicals as a result of extensive flooding. METHODS: The survey consisted of an environmental exposure assessment, which included assaying water and soil samples for contaminants, and taking blood and urine samples from 45 adolescents aged 15-18 years. We also made a subjective questionnaire assessment of 155 households. FINDINGS: There was significant contamination of the soil in Istoca, but no water contamination in the aftermath of hurricane Mitch. The soil levels of chlopyrifos and parathion were 30- and 1000-times higher, respectively, than the Environmental Data Quality Level. However, the most striking finding was the detection of elevated levels of chlorinated and organophosphate pesticides in adolescents. Toxicological analyses of serum specimens showed that 51% of the samples had elevated levels of 1,1-dichloro-2,2-bis-(p-chlorophenyl) ethylene (p,p-DDE) (range, 1.16-96.9 ng/ml) (US reference mean = 3.5 ng/ml) in adults). Dieldrin levels > 0.2 ng/ml were also present in 23% of the serum specimens (serum levels of this analyte in US adolescents are < 0.2 ng/ml). Of 43 urine samples analysed for organophosphate metabolites, 18.6% contained diethyl phosphate (DEP) at levels which were greater that the reference mean of 6.45 micrograms/g creatinine. We also detected elevated levels of p-nitrophenol (p-NP) and of 3,5,6-trichloro-2-pyridinol (3,5,6-TCPY) in 91% and 42% of the samples, respectively. CONCLUSIONS: The elevated levels of chlorinated pesticides were surprising, since although these substances were banned in Honduras 15 years ago it appears that they are still being used in the country. Moreover, elevated levels of organophosphates were detected in the study adolescents even three weeks after the hurricane. Since these chemicals are usually cleared from the body quickly, our data suggest that the adolescents face an ongoing threat from pesticide exposure.

Teaching future professors how to teach.

This paper describes a course designed to provide hands-on teaching experience to future professors and to incorporate techniques for more effective teaching. A team of Ph.D. candidates, under the direction of a senior faculty member, prepared a new course from beginning to end and then offered it to a class of graduate students. The course was developed using the unit map concept so that the presentations by the five student-instructors complemented and built upon one another. Immediately after each class, feedback was given to the student-instructors by the faculty advisor and the other student-instructors. Review of video tapes of the lecture reinforced this feedback. At the completion of the course, both students and student-instructors were surveyed as to the effectiveness of the course and the student-instructors. This teaching experience and the feedback obtained from the surveys will be invaluable to the student-instructors in their future development.

Delineation of key amino acid side chains and peptide domains for antimicrobial properties of divercin V41, a pediocin-like bacteriocin secreted by Carnobacterium divergens V41.

Divercin V41 (DV41) is a class IIa bacteriocin produced by Carnobacterium divergens V41. This antilisterial peptide is homologous to pediocin PA-1 and contains two disulfide bonds. To establish the structure-activity relationships of this specific family of bacteriocin, chemical modifications and enzymatic hydrolysis were performed on DV41. Alteration of the net charge of this cationic bacteriocin by succinylation and acetylation revealed that, in a certain range, the electrostatic interactions were surprisingly not necessary for the activity of DV41. Cleavage of DV41 by endoproteinase Asp-N released two fragments N1[1-17] and N2[18-43] corresponding to the conserved hydrophilic N-terminal and the variable hydrophobic C-terminal sequences, respectively. Inhibitory assays showed that only the C-terminal fragment was active, and after trypsin cleavage at Lys42 or disulfide reduction it lost its inhibitory activity. These results suggested that both hydrophobicity and folding imposed by the Cys25-Cys43 disulfide bond were essential for antilisterial activity of the C-terminal hydrophobic peptide. Chemical oxidation of tryptophan residues by N-bromosuccinimide demonstrated that these residues were crucial for inhibitory activity since modification of any one of them rendered DV41 inactive. On the contrary, only the modification of all the three tyrosine residues caused a total loss of antilisterial activity. These latter results strengthened previous results suggesting that the N-terminal domain containing the YGNGV consensus sequence was not involved in the binding of DV41 to a potential specific receptor on listerial cells.

Utility of serum progesterone and prolactin analysis for assessing reproductive status in the Asian elephant (Elephas maximus).

Concentrations of serum progesterone and prolactin were assessed during the perioestrous period and throughout gestation in the Asian elephant (Elephas maximus) as a means of generating information of potential use to managers. In > 95% of perioestrous periods (n=35), behavioural oestrus (as determined by bull interest, mounting and/or breeding) coincided with the onset of increased serum progesterone concentrations at the beginning of the luteal phase and continued through Day 7 (Day 1 = first significant serum progesterone rise). Within individuals, 1- to 2-day transient decreases (P < 0.05) in serum progesterone occurred between Days 2 and 9. Notably, no sexual behaviour was observed in any female after this transient fall in progesterone. Prolactin concentrations fluctuated randomly throughout the perioestrous period, with no clear pattern. During the study, four females conceived (one conceived twice), and two delivered three viable offspring. Serum progesterone was elevated above baseline throughout gestation, and then declined precipitously 2-3 days before parturition. Serum prolactin concentrations were significantly elevated above baseline (P < 0.05) after 5-6 months of gestation and remained high until after parturition. This study confirms that serum progesterone and prolactin analyses are useful tools for monitoring the reproductive status of Asian elephant females. Specifically, the transition from low to high progesterone secretion during the late interluteal/early luteal phase is predictive of oestrus and can be used to coordinate breeding efforts. Pregnancy can be confirmed by elevated serum prolactin after 6 months postbreeding, whereas the late gestational decrease in progesterone is predictive of impending parturition.

Improvement of coronary vasodilatation capacity through single LDL apheresis.

A concomitant phenomenon of hypercholesterolemia is reduced coronary vasodilatation capacity due to disturbed endothelial function. Endothelial function can be partially or completely normalized by reducing cholesterol levels through drug therapy, but it is still unclear how rapidly this desired effect is achieved. An interval of between weeks and months has been presumed. LDL apheresis (LDL-A) is capable of achieving a high-degree LDL cholesterol reduction within hours. With positron emission tomography (PET), carried out immediately before and after LDL-A, changes in coronary reserve due to this abrupt LDL cholesterol reduction could be measured both quantitatively and non-invasively. In nine patients (six women, three men) with documented coronary artery disease and hypercholesterolemia, PET was carried out immediately before and 18-20 h after LDL-A. A reduction in LDL cholesterol (from 194 +/- 38 to 81 +/- 20 mg/dl), facilitated significant improvement in myocardial blood flow (MBF) (173 +/- 63 versus 226 +/- 79 ml/min per 100 g) after pharmacologic recruitment of coronary flow capacity (dipyridamole stress), coronary flow reserve (CFR) (1.91 +/- 0.68 versus 2.48 +/- 0.68) and minimum coronary resistance (MCR) (0.61 +/- 0.18 versus 0.43 +/- 0.16 mmHg/100 g per min per ml) within 24 h. Plasma viscosity was reduced slightly, by 6.6%. Probably for the first time, a 30% improvement in coronary vasodilatation capacity could be demonstrated quantitatively and non-invasively by PET after a single LDL-A within 24 h.

Microbial characterization of biological filters used for drinking water treatment.

The impact of preozonation and filter contact time (depth) on microbial communities was examined in drinking water biofilters treating Ohio River water which had undergone conventional treatment (coagulation, flocculation, sedimentation) or solutions of natural organic matter isolated from groundwater (both ozonated and nonozonated). With respect to filter depth, compared to filters treating nonozonated waters, preozonation of treated water led to greater differences in community phospholipid fatty acid (PLFA) profiles, utilization of sole carbon sources (Biolog), and arbitrarily primed PCR fingerprints. PLFA profiles indicated that there was a shift toward anaerobic bacteria in the communities found in the filter treating ozonated water compared to the communities found in the filter treating nonozonated settled water, which had a greater abundance of eukaryotic markers.

Properties of low density lipoproteins relevant to oxidative modifications change paradoxically during aging.

Atherosclerosis is a common problem among the elderly. Because lipid peroxidation is considered a contributor to the development of atherosclerosis, we compared oxidative properties of lipoproteins in an otherwise healthy (SENIEUR-classified) aged population (65-74 years) with young controls (18-30 years). Relative amounts of oxidatively altered low density lipoprotein (LDL), estimated by means of an antibody against LDL modified by 4-hydroxynonenal, a product of lipid peroxidation, were increased marginally in serum from the elderly (9.8 vs. 7.4%, P = 0.07). In contrast, isolated LDL from the elderly revealed a decreased susceptibility to in vitro oxidation: the lag time was increased (2.34 vs. 2.10 h, P < 0.01), and the maximal rate of LDL oxidation decreased (0.88 vs. 1.01 O.D./h, P = 0.001). However, there were no age-related changes in lipid composition of native LDL and consumption of fatty acids during in vitro oxidation. The serum concentrations of ascorbic acid and most lipophilic anti-oxidants (the latter expressed per g serum lipids) were significantly decreased in the elderly except tocopherols which tended to be higher. In conclusion, our data reveal paradox age-related alterations of LDL as to its behaviour in oxidation in vivo vs. in vitro.

[The antihypertensive effect of the new cardioselective prolonged-action beta-adrenoblocker bisoprolol compared with propranolol, metoprolol and placebo]. / Antigipertenzivnyi éffekt novogo kardioselektivnogo beta-adrenoblokatora prolongirovannogo deistviia bisoprolola v sravnenii s propranololom, metoprololom i platsebo.

The antihypertensive effect of daily doses of three beta-adrenoblockers (Bisoprolol, 10 mg once a day, propranolol, 80 mg twice a day, and methoprolol, 100 mg twice a day), and placebo was examined in 14 patients with persistent mild and moderate hypertension during a double blind cross-over study by using 24-hour monitoring of blood pressure and its routine measurements. The latter made by a mercury sphygmomanometer indicated that the antihypertensive and negative chronotropic effect of Bisoprolol in a dose of 10 mg remained 24 hours after its administration and it did not significantly differ from that of the two other agents given in the above doses. The application of 24-hour blood pressure monitoring allows a more pronounced antihypertensive effect of bisoprolol to be revealed during 24 hours than that displayed by the two agents. Bisoprolol is an effective and safe antihypertensive agent.

Low density lipoprotein for oxidation and metabolic studies. Isolation from small volumes of plasma using a tabletop ultracentrifuge.

A rapid method is described for the isolation of small volumes of plasma low density lipoprotein (LDL) free of plasma protein contaminants using the TL-100 Tabletop Ultracentrifuge (Beckman). The isolation of LDL was achieved by a 25 min discontinuous gradient density centrifugation between the density range of 1.006 and 1.21 g/ml, recovery of LDL by tube slicing followed by a 90 min flotation step (d = 1.12 g/ml). The purity of LDL and apolipoprotein B100 (apo B100) were monitored by agarose electrophoresis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), radial immunodiffusion and micropreparative fast protein liquid chromatography (FPLC). The ability of LDL oxidation was assessed by following absorbance at 234 nm after addition of copper ions. The functional integrity of the isolated LDL was checked by clearance kinetics after injection of [125I]-labelled LDL in estrogen-treated rats. The additional purification step led to LDL fractions free of protein contamination and left apo B100, alpha-tocopherol and beta-carotene intact. The LDL prepared in this way was free of albumin, as evident from analytic tests and from its enhanced oxidative modification by copper ions. Used for analytical purposes, this method allows LDL preparations from plasma volumes up to 570 microliters. This method is also convenient for metabolic studies in small animals, especially those relating to the determination of kinetic parameters of LDL in which LDL-apo B100 has to be specifically radiolabelled.

Seroprevalence of hepatitis A, B, C, and D markers and liver function abnormalities in intravenous heroin addicts.

To determine the most prevalent forms of hepatitis in intravenous heroin addicts, 389 addicts consecutively admitted to outpatient treatment clinics throughout California were tested for antibodies to hepatitis A (anti-HAV), B core (anti-HBc), B surface (anti-HBs), C (anti-HCV), D (anti-HDV), and B surface antigen (HBsAg). The majority were also tested for serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase, lactic dehydrogenase, total bilirubin, globulins, albumin, and platelet count. The seroprevalence of each marker was: anti-HAV (40.7%); anti-Hbc (73.6%); anti-HBs (46.7%); anti-HCV (93.6%); anti-HDV (9.6%), and HBsAg (3.5%). No single case was positive for IgM, anti-HAV, or for both HBsAg and anti-HDV, indicating the presence of recent hepatitis A or hepatitis D infection. Abnormal liver enzymes, serum proteins, total bilirubin, and platelet count were found to be normal in 5.3 to 44.8% of anti-HCV cases indicating persistent infection. Among anti-HCV cases, elevated total bilirubin or a low platelet count was invariably associated with one or more liver enzyme and protein abnormalities. We conclude that while acute hepatitis may be frequent and caused by various viral types, hepatitis C is the primary form of chronic hepatitis found in intravenous heroin addicts. Almost half of hepatitis C cases demonstrate liver function abnormalities indicating persistent infection that has the potential to be contagious and progress to cirrhosis, liver failure, and hepatocellular carcinoma.

[Metal concentrations in benthic marine algae in 3 regions of the state of Rio de Janeiro]. / Concentrações de metais em algas marinhas bentônicas de três regiões do estado do Rio de Janeiro.

The concentrations of zinc, cadmium and copper were determined in different benthic algal species from some sites located in Guanabara Bay, Sepetiba Bay and Ribeira Bay (State of Rio de Janeiro). The aim is to verify the possibility of using algae as indicators of metallic contamination in these areas. According to our results, the highest concentrations of Zn and Cd in algae were observed in Sepetiba Bay. In this region, the differences among the concentrations of the metals were analysed in three sampling sites. Padina gymnospora was the species recommended to monitor the contamination of Zn and Cd in Sepetiba Bay, for the following reasons: 1) its great abundance and distribution in the areas, and 2) its high ability to accumulate metals. More studies are suggested to evaluate the levels of metals in Guanabara Bay.

Survival of microorganisms in smectite clays: implications for Martian exobiology.

Manned exploration of Mars may result in the contamination of that planet with terrestrial microbes, a situation requiring assessment of the survival potential of possible contaminating organisms. In this study, the survival of Bacillius subtilis, Azotobacter chroococcum, and the enteric bacteriophage MS2 was examined in clays representing terrestrial (Wyoming type montmorillonite) or Martian (Fe(3+)-montmorillonite) soils exposed to terrestrial and Martian environmental conditions of temperature and atmospheric pressure and composition, but not to UV flux or oxidizing conditions. Survival of bacteria was determined by standard plate counts and biochemical and physiological measurements over 112 days. Extractable lipid phosphate was used to measure microbial biomass, and the rate of 14C-acetate incorporation into microbial lipids was used to determine physiological activity. MS2 survival was assayed by plaque counts. Both bacterial types survived terrestrial or Martian conditions in Wyoming montmorillonite better than Martian conditions in Fe(3+)-montmorillonite. Decreased survival may have been caused by the lower pH of the Fe(3+)-montmorillonite compared to Wyoming montmorillonite. MS2 survived simulated Mars conditions better than the terrestrial environment, likely due to stabilization of the virus caused by the cold and dry conditions of the simulated Martian environment. The survival of MS2 in the simulated Martian environment is the first published indication that viruses may be able to survive in Martian type soils. This work may have implications for planetary protection for future Mars missions.

Three years of experience with bisoprolol in the treatment of mild to moderate hypertension.

Long-term efficacy and safety of the beta1-selective adrenoceptor-blocking agent bisoprolol was investigated in 164 patients with mild to moderate essential hypertension who had already been treated successfully with bisoprolol once daily for 1 year in preceding studies. The good therapeutic effect of bisoprolol was maintained for another 1 or 2 years. There was no need for further dose adaptations, and the patients remained on their individually titrated dose. At the end of the third year, 22.6% of the patients required 5 mg or less, 63.7% needed 10 mg, and 13.7% of the patients required 20 mg. Four patients received a diuretic in addition to maintain a reasonable blood pressure reduction. Thirty-four of the 164 patients experienced adverse events, which necessitated a premature study withdrawal only in 2 cases. The repeatedly checked laboratory parameters did not adversely change under long-term treatment with bisoprolol up to 36 months. In particular, this also holds true for total cholesterol, triglycerides, and blood glucose.

Adrenal steroidogenesis in the spontaneously hypertensive rat (SHR).

Adrenal vein catheterizations were done in SHR and control rats at different ages during the development of hypertension. All SHR became hypertensive by 15 weeks of age. The secretion rate of aldosterone was significantly reduced at 8 weeks of age, 18-hydroxy-11-deoxycorticosterone (18-OH-DOC) at 12 weeks of age, deoxycorticosterone (DOC) at twenty weeks of age, and corticosterone (B) at 12 and 20 weeks of age. Secretion data suggest either an enzyme block, or increased conversion of known steroids to an unknown steroid product. Reduced secretion of corticosterone could explain the adrenal hyperplasia observed in SHR which may be important to the development of hypertension in these animals.