The Vaginal Microbiota and Behavioral Factors Associated With Genital Candida albicans Detection in Reproductive-Age Women.

BACKGROUND: Vulvovaginal candidiasis is commonly diagnosed and has been associated in prospective studies with the acquisition of HIV. Little data is available on how the composition of the vaginal microbiota, and other risk factors, are associated with the molecular detection of Candida albicans-a common cause of vulvovaginal candidiasis. METHODS: In a cross-sectional study, self-collected vaginal swabs were obtained from 394 nonpregnant, reproductive-age women. C. albicans was detected using polymerase chain reaction targeting C. albicans ITS1/2 region. Vaginal microbiota was characterized by 16S rRNA gene amplicon sequencing of the V3 to V4 hypervariable regions and clustered into community state types (CSTs). Multiple logistic regression identified factors associated with C. albicans detection. RESULTS: Twenty-one percent had C. albicans detected and 46% reported vaginal symptoms in the prior 60 days. There was a 2-fold increase in the odds of C. albicans if a woman was in a L. crispatus-dominated CST compared to CSTs with low-Lactobacillus levels (adjusted odds ratio, 2.05; 95% confidence interval, 0.97-4.37). History of self-treatment with antifungals, L. crispatus relative abundance, and receptive oral sex were also significantly associated with C. albicans detection. CONCLUSIONS: A L. crispatus-dominated vaginal microbiota is thought to protect women from both development of bacterial vaginosis and incidence of sexually transmitted infections; however, our data suggest that L. crispatus is associated with increased C. albicans detection. Receptive oral sex may also be a risk factor for vaginal C. albicans colonization.

Determination of the normal reference interval for anti-Müllerian hormone (AMH) in bitches and use of AMH as a potential predictor of litter size.

Anti-Müllerian hormone (AMH) is a reliable endocrine marker of ovarian reserve in many species with extensive literature in both humans and cattle. However, there are no known hormonal predictors of ovarian reserve and potential reproductive performance in the bitch. A prospective cohort study was performed involving 155 intact bitches of various ages (range 1.2-7.6 years) and breeds that were presented for routine breeding management over a one-year period. All bitches were artificially inseminated with frozen or fresh semen using the transcervical insemination (TCI) technique. AMH concentrations were measured using a commercially available canine AMH ELISA (Ansh Labs® , Texas, USA), which we validated prior to performing the study. The reference interval (RI) for AMH for all bitches in the study, regardless of body weight, was 2.9-21.1 ng/ml. There was a significant effect of bitch size and age on AMH concentrations. The RI for giant breeds was significantly (p < .01) lower (1.75-15.6 ng/ml) than small-sized (5.6-24.2 ng/ml), medium-sized (4.3-23.7 ng/ml) and large-sized (4.3-21.0 ng/ml) bitches. The mean AMH concentration in bitches less than 4 years of age was 12.4 ng/ml, whereas the mean AMH concentration in bitches older than 4 years of age was 10.5 ng/ml (p < .05). For each additional year of age above 1 year of age, AMH concentrations fell by 0.5 ng/ml. There was no effect of AMH concentration on the whelping rate. Smaller breeds had smaller litters (and higher AMH concentrations), but within each breed size category, bitches with higher AMH concentrations had significantly larger litter sizes (p < .01). For each 1 ng/ml increase in AMH, litter size increased by 0.3 pups/litter. In conclusion, we determined a normal reference interval for AMH in bitches based on body weight using a canine-specific assay. In agreement with findings in humans and other species, we found that there is a decline in AMH concentrations with advancing age in bitches. Finally, the significant positive correlation between AMH concentrations and litter size indicates that AMH may be a useful management tool for the selection of bitches in breeding programmes.

Periparturient characteristics of mares and their foals on a New Zealand Thoroughbred stud farm.

AIMS: To describe selected periparturient variables in a sample of Thoroughbred mares and their foals on a commercial stud farm in New Zealand. METHODS: Data were collected for 969 foaling records, from 592 mares, by foaling attendants from one Thoroughbred stud farm in the Waikato region of New Zealand for the breeding seasons 2008-2013, inclusive, and collated retrospectively. The associations between mare- and foal-level variables and gestation length, time of birth (expressed as the number of hours after midday), fetal membrane retention time, time for the foal to stand and suckle after birth, and foal mortality were quantified using linear and logistic regression models. RESULTS: Mean gestation length for mares in this study was 350 (min 296, max 429) days, and was shorter for filly foals (348 (min 296, max 429) days) compared with colt foals (350 (min 310, max 404) days) (p=0.001). Overall, 517/968 (53%) foalings occurred between 19:00 and 01:00 and 212/968 (22%) foalings occurred during daylight. Overall, 815/855 (95%) expelled their fetal membranes within 4 hours of parturition. The mean interval from birth to standing and to suckling was shorter (p<0.001) for filly foals than colts by 13 (95% CI=9-17) and 17 (95% CI=8-26) minutes, respectively. Dystocia was reported for 81/968 (8.4%) foalings with relative fetal oversize being the most common cause (36/81 (44%) cases). Perinatal foal death was recorded for 26/966 (2.7%) foalings. The odds of foal mortality decreased with increasing length of gestation (p=0.027), and were greater for colt than filly foals (p=0.042) and when dystocia was reported (p<0.001). CONCLUSIONS: Gestation length was influenced by the sex of the foal, as was the time taken for the foal to stand and suckle after birth. One fifth of mares foaled during daylight hours. The majority of mares in this study population expelled their fetal membranes within 4 hours of foaling. CLINICAL RELEVANCE: Accurate measures for periparturient behaviour can be important in the management of mares and newborn foals. Clinically relevant differences appear to exist between mares and foals in New Zealand and those in the Northern Hemisphere for some key parameters, likely as a result of differences in management and environment. The results of this study suggest that mares should be considered to have retained their membranes if not passed within 4 hours of parturition.

Revealing the nature of excitons in liquid exfoliated monolayer tungsten disulphide.

Transition metal dichalcogenides (TMD) hold promise for applications in novel optoelectronic devices. There is therefore a need for materials that can be obtained in large quantities and with well understood optical properties. In this report, we present thorough photoluminescence (PL) investigations of monolayer tungsten disulphide obtained via liquid phase exfoliation. As shown by microscopy studies, the exfoliated nanosheets have dimensions of tens of nanometers and thickness of 2.5 monolayers on average. The monolayer content is about 20%. Our studies show that at low temperature the PL is dominated by excitons localized on nanosheet edges. As a consequence, the PL is strongly sensitive to the environment and exhibits an enhanced splitting in magnetic field. As the temperature is increased, the excitons are thermally excited out of the defect states and the dominant transition is that of the negatively charged exciton. Furthermore, upon excitation with a circularly polarized light, the PL retains a degree of polarization reaching 50% and inherited from the valley polarized photoexcited excitons. The studies of PL dynamics reveal that the PL lifetime is on the order of 10 ps, which is probably limited by non-radiative processes. Our results underline the potential of liquid exfoliated TMD monolayers in large scale optoelectronic devices.

Use of equine embryo -derived mesenchymal stromal cells and their extracellular vesicles as a treatment for persistent breeding-induced endometritis in susceptible mares.

Persistent breeding induced endometritis (PBIE) is a significant cause of infertility in mares. The development of a safe, universal, readily available therapeutic to manage PBIE and facilitate an optimal uterine environment for embryo development may improve pregnancy rates in susceptible mares. Mesenchymal stromal cells (MSCs) are being used increasingly as a therapeutic mediator for inflammatory conditions such as endometritis, and early gestational tissue provides a unique source of multipotent stem cells for creating MSCs. Extracellular vesicles (EVs) are mediators of cell communication produced by many different cell types. This study utilized embryo-derived mesenchymal stromal cells (EDMSCs) and their EVs as a potential therapeutic modality for PBIE in two groups: a) PBIE-susceptible mares challenged with pooled dead sperm (n=5); and b) client-owned mares diagnosed as susceptible to PBIE (n=37 mares and 40 estrous cycles). Mares pre-treated with intrauterine EDMSCs or their EVs resulted in a significant reduction in the accumulation of intrauterine fluid post-breeding. Nine of 19 (47 %) mares treated with EDMSCs prior to natural breeding and 13 of 20 (65 %) mares treated with EDMSC derived EVs were pregnant after the first cycle and 12 of 18 (67 %) mares treated with EDMSCs, and 15 of 19 (79 %) mares treated with EVs conceived by the end of the breeding season. These preliminary clinical studies are the first reports of the use of EDMSCs or their EVs as a potential intrauterine therapy for the management of PBIE susceptible mares.

Observational cohort study of the effect of a single lubricant exposure during transvaginal ultrasound on cell-shedding from the vaginal epithelium.

The outer layers of the vaginal epithelium (VE) are important because they accumulate glycogen which, under optimal conditions, Lactobacillus spp. consume to grow and acidify the vaginal microenvironment with lactic acid. We hypothesized that exposure to lubricant, for example in the conduct of a transvaginal ultrasound (TVUS), may contribute to the shedding of mature epithelial cells, exposing immature cells. Cervicovaginal fluid (CVF) was sampled at four time points by menstrual cup (Softdisc™) from 50 women referred for TVUS, during which a controlled volume of lubricant was applied to the TVUS wand. Samples were collected (1) immediately before TVUS and (2) 6-12 hours, (3) within one week, and (4) two weeks after TVUS. Clinical vaginal lubricants are similar to commercial lubricants, and often have a high osmolality or pH, and contain bactericides such as methylparaben and propylparaben. The number and maturity of epithelial cells in each CVF sample were measured by quantitative and differential fluorimetry (maturity index, MI). Comparisons of cell-counts and maturity were made by paired Wilcoxon signed-rank tests. Among women with a high pre-TVUS MI (> 3), there was a decrease in median cell-count and mean MI in the sample collected 6-12 hours after TVUS (p<0.001, n = 26 and p < 0.001, n = 26, respectively). For these women, cell-count and MI remained lower in the sample collected within the subsequent week (p<0.001, n = 29 and p<0.01, n = 29, respectively), and MI remained lower in the sample collected within two weeks of TVUS (p<0.01, n = 25), compared to the pre-TVUS sample. Among participants with a low pre-TVUS MI (< 3), cell-count was higher in the sample collected within two weeks of TVUS compared to the pre-TVUS sample (p = 0.03, n = 15), but no significant changes in MI were observed. Results were similar when restricted to reproductive-age women. This preliminary data indicates hypertonic vaginal lubricants may increase vaginal epithelial cell shedding.

Factors influencing parentage ratio in canine dual-sired litters.

Breeding a bitch with two different sires during a single estrous cycle has the potential to facilitate rapid genetic gain and improve reproductive performance within a canine breeding colony. There is limited data regarding the factors that contribute to the success of dual-sired litters in domestic dogs, and only anecdotal evidence suggesting that these litters rarely produce offspring from more than one sire. The objective of this prospective clinical study was to investigate multiple factors that likely affect the success of dual-sired litters on whelping rate, litter size and parentage ratio. These factors include: timing of artificial insemination (AI), order of sires, number of AI's per cycle, semen type, sperm quality and age of sire and bitch. Data collected over a 10 year period from twenty-nine estrous cycles (28 individual bitches of 10 different breeds) were evaluated after an initial AI with frozen semen from the 'genetically desired' sire and followed up with a second AI with either fresh (n = 9) or frozen (n = 16) semen or natural mating (n = 4) from a different, 'back up', sire. DNA parentage of each pup born was determined by using a primary panel of 288 SNPs. The whelping rate and litter size from previous single sire inseminations per estrous cycle, in the same bitches, (n = 16) over 25 estrous cycles using either fresh (n = 4) or frozen-thawed (n = 21) semen, were analyzed as controls. Of the 29 dual-sired breedings, 26 bitches whelped (89.7%), and 8 litters (30.8%) were of mixed parentage. In the litters of mixed parentage after a dual-sired breeding, a greater proportion of the offspring were from the second sire than the first sire (73.0% and 27.0% respectively; P < 0.05). Interestingly, in litters where all pups were of single sire parentage after a dual-sired breeding, 50.0% of the offspring were by the first sire and 50.0% were by the second sire. For litters of mixed or single paternity produced by dual-sired breeding there was no difference in average litter size. However, on a per estrous cycle basis for each bitch the whelping rate (89.7% v. 76.0%.) and litter size (5.5 ± 2.5 v. 4.0 ± 2.78) of all dual-sired breedings were greater (P < 0.05) compared to previous single-sired breedings (controls) respectively. This study demonstrates that offspring of mixed parentage derived from dual-sired breedings may be achieved. Furthermore, insemination with semen from two different sires may increase the whelping rate and litter size, which is an important consideration when using genetically valuable, or older individuals with potentially reduced fertility.

Asymptomatic Bacterial Vaginosis Is Associated With Depletion of Mature Superficial Cells Shed From the Vaginal Epithelium.

Previous studies have described bacterial vaginosis (BV) as associated with increased cell-shedding from the cervicovaginal epithelium. Cell-shedding in excess of cell-proliferation is thought to decrease epithelial barrier function and increase susceptibility to infection. This study evaluated the number of shed cells in mid-vaginal smears from women with a diagnosis of symptomatic BV (sBV, n = 17), asymptomatic BV (aBV, n = 71), or no BV (n = 104) by Amsel criteria. The sBV smears contained significantly more shed cells (median 158/100X field) than no BV smears (median 91/100X field), p = 7.2e-9. However, we observed that aBV smears contained significantly fewer shed cells (median 35/100X field) than no BV smears, p = 22.0e-16. The sizes of cell-aggregates (cells shed in sometimes multilayered sections with intact cell-cell attachments) followed the same pattern. Cell-aggregates in sBV smears were significantly larger (median ~220,000 µm2) than those in no BV smears (median ~50,000 µm2), p = 1.8e-6, but cell-aggregates in aBV smears were significantly smaller (median ~7,000 µm2) than those in no BV smears, p = 0.0028. We also compared the superficial cell index (SCI), a measure of cervicovaginal epithelial cell maturity, in no BV and aBV smears with relatively low numbers of shed cells (≤50/100X field). The SCI of no BV smears was significantly higher (median 0.86) than that of aBV smears (median 0.35), p = 4.3e-98, suggesting a depletion of mature cells with exposure and shedding of underlying immature cells in aBV with low number of shed cells. These results indicate that aBV may contribute disproportionately to the increased susceptibility to reproductive tract infections associated with BV. Our findings remained true when considering only those smears in which the microbiota comprised a diverse set of strict and facultative anaerobic bacteria [Community State Type IV (n = 162)], thus excluding those dominated by Lactobacillus spp. This is consistent with our developing hypothesis that high-shedding sBV and low-shedding aBV could be temporally separated phases of the same condition, rather than two separate forms of BV. These findings might inform future work on clinical management of symptomatic and asymptomatic bacterial vaginosis.

Gene encoding the beta subunit of S100 protein is on chromosome 21: implications for Down syndrome.

S100 protein is a calcium-binding protein found predominantly in the vertebrate nervous system. Genomic and complementary DNA probes were used in conjunction with a panel of rodent-human somatic cell hybrids to assign the gene for the beta subunit of S100 protein to the distal half of the long arm of human chromosome 21. This gene was identified as a candidate sequence which, when expressed in the trisomic state, may underlie the neurologic disturbances in Down syndrome.

Characterization of KIFC2, a neuronal kinesin superfamily member in mouse.

Members of the kinesin superfamily of microtubule-associated proteins are involved in a variety of intracellular processes including cell division and organelle transport. In the case of axonal transport, all kinesin superfamily members reported thus far appear to play a role in anterograde transport, while a different type of microtubule motor, dynein, appears to function in retrograde transport. To better understand the role of kinesins in axonal transport, we cloned and characterized KIFC2, a novel kinesin superfamily member from mouse brain. KIFC2 encodes a 792 amino acid protein, which contains the conserved motor domain at the C-terminal end of the protein and is most similar to members of the KAR3 family involved in cell division. However, expression analysis localized KIFC2 mRNA to nonproliferative neuronal cells in the central nervous system, and immunolocalization studies demonstrated that KIFC2 is present in axons and dendrites of neurons in the central and peripheral nervous systems. Immunolocalization and biochemical fractionation studies suggest that KIFC2 localizes with some, but not all, axonally transported organelles. Finally, ligation of mouse peripheral nerves showed that KIFC2 accumulates at the proximal and distal sides of an axonal ligature. Taken together, the data suggest that, unlike other C-terminal motor proteins that appear to be involved in cell division, KIFC2 may play a role in retrograde axonal transport.

Inhibition of norepinephrine-induced cardiac hypertrophy in s100beta transgenic mice.

We have recently reported that the Ca2+-binding protein S100beta was induced in rat heart after infarction and forced expression of S100beta in neonatal rat cardiac myocyte cultures inhibited alpha1-adrenergic induction of beta myosin heavy chain (MHC) and skeletal alpha-actin (skACT). We now extend this work by showing that S100beta is induced in hearts of human subjects after myocardial infarction. Furthermore, to determine whether overexpression of S100beta was sufficient to inhibit in vivo hypertrophy, transgenic mice containing multiple copies of the human gene under the control of its own promoter, and CD1 control mice were treated with norepinephrine (NE) (1.5 mg/kg) or vehicle, intraperitoneally twice daily for 15 d. In CD1, NE produced an increase in left ventricular/body weight ratio, ventricular wall thickness, induction of skACT, atrial natriuretic factor, betaMHC, and downregulation of alphaMHC. In transgenic mice, NE induced S100beta transgene mRNA and protein, but provoked neither hypertrophy nor regulated cardiac-specific gene expression. NE induced hypertrophy in cultured CD1 but not S100beta transgenic myocytes, confirming that the effects of S100beta on cardiac mass reflected myocyte-specific responses. These transgenic studies complement in vitro data and support the hypothesis that S100beta acts as an intrinsic negative regulator of the myocardial hypertrophic response.

Factors affecting the reproductive performance of bitches: A prospective cohort study involving 1203 inseminations with fresh and frozen semen.

The aim of this prospective cohort study was to utilize multivariable statistical methods to identify factors that significantly affected whelping rate, litter size and gestation length in a large population of bitches of many different breeds, presented for routine breeding management. In addition, we aimed to determine the incidence of dystocia and the proportion of bitches undergoing a caesarean section procedure. A total of 1146 individual bitches representing 84 different breeds contributed 1203 inseminations over the 9 year (2007-2015) study period. Bitches were inseminated with either frozen-thawed (n = 645), fresh (n = 543) or chilled (n = 15) semen from 1371 different males. The mean (SD) whelping rate was 74± 4% and the mean litter size was 5.8 ± 3.1 pups per litter for all bitches in the study. The whelping rate was significantly lower in bitches inseminated with frozen-thawed semen compared with bitches inseminated with fresh semen (71% vs 80% respectively; P < 0.001). Semen that was classified as having poor motility (<30% progressive) resulted in a significantly lower whelping rate (37%) than semen classified as good (30-65% progressive; whelping rate = 67%) or excellent (>65% progressive; whelping rate = 79%). There was a linear decline in whelping rate with advancing age. Greyhounds and Labradors demonstrated a significantly higher whelping rate (88% and 94% respectively) compared with all other breeds (71.3%, P < 0.001). Bitches inseminated with frozen-thawed semen had significantly smaller litter sizes than bitches inseminated with fresh semen (5.4 ± 3.1 vs 6.2 ± 3.0 pups per litter respectively; P = 0.02). Smaller breeds had significantly smaller litters (4.4 ± 2.1 pups) than medium (5.2 ± 2.9 pups), large (5.9 ± 2.9 pups) or giant (6.7 ± 3.8 pups) breeds. For each advancing year of age, litter size decreased by 0.13 pups per litter. The mean (SD) gestation length from LH0 was 65 ± 1.9 d. Greyhounds had a significantly longer pregnancy duration (68.0 ± 1.5 d) than other breeds. For each additional year of bitch age, gestation length increased by 0.11 days (P < 0.01), and for each additional pup per litter, gestation length was reduced by 0.08 days (P < 0.05). Of the 890 bitches for which whelping outcomes were recorded; 409 (46%) whelped normally without assistance, 249 (28%) had an elective C-section, 205 (23%) underwent an emergency C-section and 27 (3%) were medically managed or required veterinary assistance for dystocia. Brachycephalic breeds were 11.3 (95CI = 9.3-17.9; P < 0.001) times more likely to have a C-section compared to all other breeds. Bitches with litter sizes of one or two pups had a C-section rate of 83%, whereas bitches with litter sizes of three or more pups had a C-section rate of 43% (P < 0.001). This study provides important clinical information to optimise whelping rates, litter size and the prediction of whelping in certain breeds for clinicians working in canine reproduction.

The first reported outbreak of equine herpesvirus myeloencephalopathy in New Zealand.

CASE HISTORY AND CLINICAL FINDINGS: On 9 January 2014 (Day 0) a mare from a stud farm in the Waikato region presented with urinary incontinence without pyrexia. Over the following 33 days 15 mares were clinically affected with neurological signs. All but one mare had a foal at foot. The most commonly observed clinical signs were hind limb paresis and ataxia. In some cases recumbency occurred very early in the course of disease and seven mares were subject to euthanasia for humane reasons. LABORATORY FINDINGS: Equid herpesvirus (EHV) type 1 was detected using PCR in various tissues collected post mortem from two mares with neurological signs. DNA sequencing data from the DNA polymerase gene of the virus showed a nucleotide transition at position 2254, a mutation encoding amino acid D752 that is highly associated with the neuropathogenic genotype of EHV-1. In total 12/15 mares were confirmed positive for EHV-1 on PCR. Results from a virus neutralisation test and ELISA on paired serum samples, and PCR on whole blood and nasal swabs, indicated that of four paddocks in a high-risk area where a cluster of cases had occurred, 20/21 (95%) horses were likely to have been exposed or were confirmed infected with EHV-1. Subsequent to the outbreak two mares aborted, one at 9 months and one at 10 months of gestation. The cause of abortion was confirmed as EHV-1 with the same genotype as that involved in the outbreak. DIAGNOSIS: Equine herpesvirus myeloencephalopathy. CLINICAL RELEVANCE: The outbreak described shows the considerable impact that can occur in outbreaks of equine herpesvirus myeloencephalopathy in New Zealand. Early biosecurity controls not only reduced the effect on the farm but mitigated the potential for the virus to spread to other horse enterprises.

Purification and reconstitution of the methyl-accepting chemotaxis proteins from Bacillus subtilis.

The methyl-accepting chemotaxis proteins (MCPs) from Bacillus subtilis, designated as H1, H2, and H3, have been purified to near homogeneity. These purified MCPs were reconstituted into proteoliposome vesicles using a detergent dilution procedure. The ability of the reconstituted MCPs to be methylated in vitro strongly suggests that they are in a functionally active conformation. The MCPs of B. subtilis are considerably larger than those of Escherichia coli, with molecular weights of the purified proteins being 76, 86, and 97 kDa for H3, H2, and H1, respectively. Two-dimensional electrophoresis demonstrates that the isoelectric point of H1 and H2 is 5.1, while H3 is slightly more basic, having an isoelectric point of 5.3. Immunoblot analysis using the cross reacting E. coli anti-Trg antibody reveals that maximal MCP expression occurs approx. 4 h after the onset of stationary phase, and remains relatively stable thereafter. However, the ability of the MCPs to be methylated in vivo is significantly reduced.

Attitudes of elderly patients and their families toward physician-assisted suicide.

OBJECTIVES: To examine and compare attitudes of elderly outpatients and their families toward physician-assisted suicide (PAS), explore sociodemographic and health correlates of these attitudes, assess family members' ability to predict patients' attitudes toward PAS, and determine family members' ability to agree on these predictions. SUBJECTS AND METHODS: Elderly patients with medical and psychiatric problems (n = 168; mean age, 75.8 years) who were attending a geriatrics specialty clinic, along with accompanying family members (n = 146), were systematically surveyed on their attitudes toward PAS in case of terminal illness, chronic illness, and mental incompetence. Relatives were also asked to predict patients' responses to items on the questionnaire. Patients and relatives were blinded to each others' responses. RESULTS: Favorable attitudes toward PAS were reported by 39.9% of the patients and 59.3% of the relatives (P < .001) in case of terminal illness, 18.2% and 25.3%, respectively, in case of chronic illness, 13.5% and 15.4%, respectively, in case of mental incompetence, and 34.0% and 55.6% (P < .001), respectively, for legalization of PAS. Family members showed a marginal ability to predict patients' attitudes toward PAS with kappa values of agreement that ranged from 0.09 to 0.41. Family members also had difficulty agreeing with each other on how they thought patients would respond (range of kappa values, 0.18-0.47). Patients who opposed PAS were women, black individuals, and those with less education, low incomes, and dementia or cognitive impairment. CONCLUSIONS: While many frail elderly patients favored PAS in cases of terminal illness, the proportion that opposed it was significantly higher than that among relatives; relatives, in turn, displayed only a marginal ability either to predict patients' attitudes or to agree among themselves. Patients who oppose PAS represent a particularly vulnerable element of society (elderly persons, women, black individuals, and poor, uneducated, and demented persons), and such patients may warrant special protection.

Supplementing previously treated anestrous dairy cows with progesterone does not increase first-service conception rate.

The objective of this study was to investigate the effect of supplementing previously treated anovulatory anestrous (AA) dairy cows with progesterone delivered intra-vaginally for 7 days, commencing 4 or 5 days after insemination, on first-service conception rate. A clinical trial, involving 990 AA dairy cows in 14 dairy herds, was conducted during the 2002/2003 breeding season. On Day -8, all cows were treated with a progesterone-containing intravaginal device (Cue-Mate). The devices were removed on Day -2; on Day -1, all cows were given 1mg of estradiol benzoate im. Cows in the control group (n = 499) received no further treatments. Cows in the treatment group (n = 491) that had been inseminated on Day 0 or 1 had a new device inserted (on Day 4 or 5), with removal of the device after 7 days. First-service conception rates for the control and treatment groups were not different (35.0% versus 36.7% respectively; P = 0.41). Furthermore, there was no difference in conception rates between cows that had devices inserted on Day 4 or 5 (31.3% versus 37.2% respectively; P = 0.45). In conclusion, supplementation of previously treated AA dairy cows with an intravaginal progesterone-releasing device for 7 days (commencing 4 or 5 days after insemination) did not significantly improve first-service conception rate.

The effect of hCG administration five days after insemination on the first service conception rate of anestrous dairy cows.

The aim of this study was to investigate the effect of treating anovulatory anestrous (AA) dairy cows with 1500 IU of hCG IM, 5 d after insemination, on their first service conception rate. A clinical trial was conducted during the 2003/2004 breeding season involving 442 AA dairy cows in six herds. On Day -8, all cows were treated with a progesterone-containing intravaginal device (Cue-Mate). The devices were removed on Day -2, and on Day -1 all cows received an IM injection of 1mg of estradiol benzoate. Cows in the control group (n=220) received no further treatments. Cows in the treatment group (n=222) which had been inseminated on Days 0 or 1 were treated with 1500 IU of hCG IM 5 d after insemination. Blood was collected from 30 cows (15 in each group) on Days 5 and 12 after AI for analysis of plasma P4 concentration. There was no difference in first service conception rates between the control and treatment groups (46.3% versus 43.6%, respectively; P=0.68), despite the fact that plasma P4 concentrations were higher in the treatment group on Day 12 (4.9+/-1.3 ng/mL versus 6.2+/-2.7 ng/mL for control and treatment groups, respectively; P<0.01). In conclusion, 1500 IU of hCG 5 d after insemination did not improve first service conception rate in AA dairy cows.

Reproductive performance in anestrous dairy cows following treatment with two protocols and two doses of progesterone.

The objectives of this study were to evaluate the effect using two doses of progesterone (P4) releasing devices in two different programs on reproductive performance of anestrous dairy cows. Cows (n = 1555) not detected in estrus by 10 d before the planned start of the seasonal breeding program and in which no CL was palpable were treated with an intravaginal P4-releasing device ('Single'; approximately 1.56 g of P4) or a modified device with triple the normal P4 dose ('Triple'; approximately 4.7 g of P4). The devices were in place for either 6 d ('Short') or 8 d ('Long'), with 1mg estradiol benzoate (EB) given 24 h after device removal. The 'Long' program also included treatment with 2 mg EB at device insertion. The Long program resulted in a higher first service conception rate (RR = 1.18 (95% CI = 1.03-1.33); P = 0.02), but had no effect on the 28-d, 56-d or final pregnancy rate compared to the Short program. There were no effects of dose of P4 on any outcome. In conclusion, the Long compared to the Short program, but not the dose of P4, improved first service conception rates in anestrous cows.

Soluble adhesion molecules (E-selectin, ICAM-1 and VCAM-1) in breast carcinoma.

Adhesion molecules are important in cell-cell and cell-basement membrane interactions. They are intimately involved in inflammatory reactions and a role in tumour progression has been postulated. E-selectin, intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) play a role in cell adhesion to the vascular endothelium, and may have a role in tumour cell dissemination. Soluble forms of these molecules have been described and this study was established to examine these adhesion molecules in patients with breast carcinoma. Serum was obtained from 92 patients with breast carcinoma and 31 age-matched patients with benign breast disease. All samples were obtained prior to surgery. Soluble levels of E-selectin, ICAM-1, and VCAM-1 were significantly elevated in patients with Stage 4 disease compared with controls. (E-selectin 88.6 (47.9) versus 51.4 (18.4) ng/ml; P<0.001: ICAM-1 447 (249) versus 244 (79) ng/ml; P<0.001: VCAM-1 779 (159) versus 552 (135) ng/ml; P<0.001 results expressed on mean (SEM) SD placed above this.). The prognostic value of the adhesion molecules was examined. In patients with Stage 2 disease, elevated VCAM-1 was predictive of decreased survival, even when corrected for T and N status. Adhesion molecules are elevated in patients with advanced disease and elevation in VCAM-1 has prognostic significance in patients with breast carcinoma.

Accumulation of S100 beta mRNA and protein in cerebellum during infancy in Down syndrome and control subjects.

S100 protein is a 20 kDA calcium-binding protein that accumulates during CNS maturation in mammals. The human gene coding for the beta subunit of S100 protein (S100 beta) is located on chromosome 21, in a subtelomeric position in 21q22.3. In order to investigate the effect of trisomy 21 on S100 beta gene expression, we performed Southern, Northern and Western blot analysis on DNA, RNA and protein, respectively, extracted from the cerebellum of control and Down syndrome (DS) subjects aged 1-18 months. Southern blot analysis revealed a novel EcoRI polymorphism in the S100 beta gene in two of 15 DNA samples examined, and a 1.5 gene dosage for S100 beta in DS. Northern and Western blot analysis showed an approximately 10-fold increase in S100 beta mRNA and protein levels between 1 and 18 months. No differences in the rates of accumulation of S100 beta mRNA and protein were observed between DS and normal subjects. These results demonstrate an increase in S100 beta mRNA and protein levels during infancy indicative of postnatal astrocytic maturation and show that there is no gross deregulation in the expression of the S100 beta gene in DS as a consequence of trisomy 21.