Silencing of Apis mellifera dorsal genes reveals their role in expression of the antimicrobial peptide defensin-1.

Like all other insects, two key signalling pathways [Toll and immune deficiency (Imd)] regulate the induction of honey bee immune effectors that target microbial pathogens. Amongst these effectors are antimicrobial peptides (AMPs) that are presumed to be produced by the nuclear factors kappa B (NF-κB) Dorsal and Relish from the Toll and Imd pathways, respectively. Using in silico analysis, we previously proposed that the honey bee AMP defensin-1 was regulated by the Toll pathway, whereas hymenoptaecin was regulated by Imd and abaecin by both the Toll and Imd pathways. Here we use an RNA interference (RNAi) assay to determine the role of Dorsal in regulating abaecin and defensin-1. Honey bees have two dorsal genes (dorsal-1 and dorsal-2) and two splicing isoforms of dorsal-1 (dorsal-1A and dorsal-1B). Accordingly, we used both single and multiple (double or triple) isoform knockdown strategies to clarify the roles of dorsal proteins and their isoforms. Down-regulation of defensin-1 was observed for dorsal-1A and dorsal-2 knockdowns, but abaecin expression was not affected by dorsal RNAi. We conclude that defensin-1 is regulated by Dorsal (Toll pathway).

Possible glimpses into early speciation: the effect of ovarian fluid on sperm velocity accords with post-copulatory isolation between two guppy populations.

Identifying mechanisms of reproductive isolation is key to understanding speciation. Among the putative mechanisms underlying reproductive isolation, sperm-female interactions (post-mating-prezygotic barriers) are arguably the hardest to identify, not least because these are likely to operate at the cellular or molecular level. Yet sperm-female interactions offer great potential to prevent the transfer of genetic information between different populations at the initial stages of speciation. Here, we provide a preliminary test for the presence of a putative post-mating-prezygotic barrier operating between three populations of Trinidadian guppies (Poecilia reticulata), an internally fertilizing fish that inhabits streams with different levels of connectivity across Trinidad. We experimentally evaluate the effect of female ovarian fluid on sperm velocity (a predictor of competitive fertilization success) according to whether males and females were from the same (native) or different (foreign) populations. Our results reveal the potential for ovarian fluid to act as a post-mating-prezygotic barrier between two populations from different drainages, but also that the strength of this barrier is different among populations. This result may explain the previous finding that, in some populations, sperm from native males have precedence over foreign sperm, which could eventually lead to reproductive isolation between these populations.

Declining sexually transmitted infections among female sex workers: the results of an HIV and sexually transmitted infection prevention strategy in Honduras, 2006-08.

In 2006, we implemented an HIV and sexually transmitted infection (STI) prevention programme for female sex workers (FSWs) in three Honduran cities. All FSW attending STI clinics underwent regular examinations and STI testing. Information on condom use with different partners was collected at each visit. After three years, we detected a significant decline in the prevalence of syphilis from 2.3% at the first screening to 0.0% at the third screening (P = 0.05), and of chlamydia, from 6.1% to 3.3% (P = 0.01). No changes were observed in the prevalence of gonorrhoea or trichomoniasis. The cumulative HIV prevalence remained constant (P = 0.44). Reports of condom use with clients increased from 93.8% to 98.9% (P < 0.001). The implementation of an HIV/STI prevention programme in FSW has contributed to increases in condom use with clients and the reduction in syphilis and chlamydia prevalence. The intervention should be strengthened and considered as part of a national health policy strategy.

First isolation and characterization of Lactococcus garvieae from Brazilian Nile tilapia, Oreochromis niloticus (L.), and pintado, Pseudoplathystoma corruscans (Spix & Agassiz).

Lactococcus garvieae infection in cultured Nile tilapia, Oreochromis niloticus (L.), and pintado, Pseudoplathystoma corruscans (Spix & Agassiz), from Brazil is reported. The commercial bacterial identification system, Biolog Microlog, confirmed the identity of L. garvieae. Infectivity trials conducted in Nile tilapia using Brazilian Nile tilapia L. garvieae isolates resulted in a median lethal dose-50 of 1.4 x 10(5) colony-forming units (CFU)/fish. This is the first evidence of the presence of this pathogen from Brazilian fish. In addition, this is the first report of L. garvieae infection in either Nile tilapia or pintado. Collectively, this evidence expands the geographical range of fish hosts, number of fish hosts harbouring L. garvieae and carbon source utilization by L. garvieae fish isolates. Furthermore, the Biolog system may be an alternative technique to polymerase chain reaction for the identification of L. garvieae and discrimination between closely related bacterial species.

Caste development and reproduction: a genome-wide analysis of hallmarks of insect eusociality.

The honey bee queen and worker castes are a model system for developmental plasticity. We used established expressed sequence tag information for a Gene Ontology based annotation of genes that are differentially expressed during caste development. Metabolic regulation emerged as a major theme, with a caste-specific difference in the expression of oxidoreductases vs. hydrolases. Motif searches in upstream regions revealed group-specific motifs, providing an entry point to cis-regulatory network studies on caste genes. For genes putatively involved in reproduction, meiosis-associated factors came out as highly conserved, whereas some determinants of embryonic axes either do not have clear orthologs (bag of marbles, gurken, torso), or appear to be lacking (trunk) in the bee genome. Our results are the outcome of a first genome-based initiative to provide an annotated framework for trends in gene regulation during female caste differentiation (representing developmental plasticity) and reproduction.

Isolation and characterization of strains of Flavobacterium columnare from Brazil.

Flavobacterium columnare is an important pathogen of freshwater fish, implicated in skin and gill disease, often causing high mortality. An outbreak of skin disease in fingerling and adult Nile tilapia, Oreochromis niloticus (L.), cultivated in a recirculation system, was investigated. Four strains were isolated and characterized by biochemical reactions, enzyme production, fatty acid profile and analysis of the 16S-23S rDNA intergenic spacer region. All strains were identified as F. columnare. Experimental infection assays with one of these strains (BZ-5-02) were conducted and pathogenicity (by intramuscular route) was demonstrated in Nile tilapia and channel catfish, Ictalurus punctatus (Rafinesque). This is the first report of characterization of Brazilian strains of F. columnare.

Additive genetic relationships between heifer pregnancy and scrotal circumference in Nellore cattle.

To estimate heritability (h2) for yearling heifer pregnancy and to estimate the genetic correlation between heifer pregnancy and scrotal circumference, 18,145 records of Nellore heifers exposed to breeding at an age of approximately 14 mo and 25,466 records of contemporary young bulls were analyzed. Heifer pregnancy was considered as a categorical trait, with the value 1 (success) assigned to heifers that were pregnant after rectal palpation approximately 60 d after the end of a 90-d breeding season and the value 0 (failure) otherwise. A single-trait animal model for heifer pregnancy and a two-trait animal model including heifer pregnancy and scrotal circumference were used. Contemporary groups were defined in two ways: including (CG2) or not including (CG1) weaning management of the heifer. Heritability estimates obtained by Method R in single-trait analyses were 0.68 +/- 0.09 and 0.61 +/- 0.10 using CG1 and CG2 definitions, respectively. Heritability estimates for two-trait analyses were 0.69 +/- 0.09 (CG1) and 0.63 +/- 0.08 (CG2) for heifer pregnancy and 0.57 +/- 0.03 (both CG) for scrotal circumference. The genetic correlation estimates between the two traits were 0.20 +/- 0.12 (CG1) and 0.20 +/- 0.13 (CG2). Based on the results of this study, EPD for heifer pregnancy can be used to select bulls for the production of precocious daughters and will be more effective than selecting on scrotal circumference EPD in Nellore cattle. However, scrotal circumference can be incorporated in a two-trait analysis to increase the accuracy of prediction for heifer pregnancy EPD for young bulls. Using contemporary group without heifer weaning management gave higher h2 and, for two-trait analysis, converged more quickly.

Vitamin D deficiency in pregnant women from a non-European ethnic minority population--an interventional study.

OBJECTIVE: To determine the vitamin D status of pregnant women from non-European ethnic minorities in South Wales. DESIGN: Prospective study. SETTING: Llandough Hospital, Cardiff, South Wales. SAMPLE: One hundred and sixty pregnant women from a non-European ethnic minority population in South Wales. METHODS: Biochemical screening of vitamin D status was carried out at the first antenatal visit. Women found to be deficient in vitamin D were subsequently supplemented and vitamin D status was rechecked at delivery. MAIN OUTCOME MEASURE: Vitamin D status at delivery. RESULTS: Eighty of 160 women had a vitamin D level below 8 ng/mL at their first antenatal visit and were treated with oral vitamin D. Factors that could influence vitamin D status such as religion, fluency in English and dressing habits did not appear to have any effect, although a higher proportion of women who had lived in Britain for longer than three years had subnormal vitamin D levels. In 58 of those checked at delivery, the mean plasma vitamin D level increased from 6 to 11 ng/mL although the mean parathyroid hormone level was unchanged. CONCLUSION: In view of the high incidence of subnormal vitamin D levels in women from ethnic minorities, we recommend biochemical screening of these women in early pregnancy, with subsequent supplementation where indicated.

Genetic evaluation of the probability of pregnancy at 14 months for Nellore heifers.

To estimate the heritability for the probability that yearling heifers would become pregnant, we analyzed the records of 11,487 Nellore animals that participated in breeding seasons at three farms in the Brazilian states of São Paulo and Mato Grosso do Sul. All heifers were exposed to a bull at the age of about 14 mo. The probability of pregnancy was analyzed as a categorical trait, with a value of 1 (success) assigned to heifers that were diagnosed pregnant by rectal palpation about 60 d after the end of the breeding season of 90 d and a value of 0 (failure) assigned to those that were not pregnant at that time. The estimate of heritability, obtained by Method R, was 0.57 with standard error of 0.01. The EPD was predicted using a maximum a posteriori threshold method and was expressed as deviations from 50% probability. The range in EPD was -24.50 to 24.55%, with a mean of 0.78% and a SD of 7.46%. We conclude that EPD for probability of pregnancy can be used to select heifers with a higher probability of being fertile. However, it is mainly recommended for the selection of bulls for the production of precocious daughters because the accuracy of prediction is higher for bulls, depending on their number of daughters.

Total body water measurement using bioelectrical impedance analysis, isotope dilution and total body potassium: a scoring system to facilitate intercomparison.

OBJECTIVES: (1) To develop a scale that is useful in evaluating the accuracy of multifrequency bioelectrical impedance analysis (MF-BIA) in the assessment of body water volumes against the accepted gold standard measurements based on isotope-dilution and total body potassium (TBK). (2) To perform a pilot test of the scale. DESIGN: A scale was developed to evaluate the accuracy of MF-BIA in the assessment of body water volumes. Questions were obtained from reading the scientific literature and discussions involving the four authors. Three of these and two additional independent readers pre-tested the scale. A weighting was identified for each question and a pilot test with a sample of 10 articles (different to those used for the questionnaire performance) was conducted. A further validation was carried out with a second set of 20 articles and two additional independent readers. RESULTS: The kappa statistic expressing the level of agreement between pairs of the first three authors using this scale with 10 articles, was 0.3, 0.4 and 0.6 after the first attempt. A second evaluation after specific changes improved the agreement to 0.8, 0.6 and 0.8. The mean score for 10 articles was 252+/-36 points from a total score of 400 (63+/-9%). The evaluation with the second set of 20 articles resulted in a kappa of 0.7 from two pairs of authors. The evaluation with two additional reviewers resulted in a kappa=0.7. CONCLUSION: A tool has been developed to assess the accuracy of the MF-BIA technique and to identify methodological components, plan future studies and critically evaluate data in this area. It is likely that this tool may also be used to assess the accuracy of single frequency studies.

Patterns of sperm precedence and predictors of paternity in the Trinidadian guppy.

Despite its widespread occurrence in animals, sperm competition has been studied in a limited range of taxa. Among the most neglected groups in this respect are internally fertilizing fish in which virtually nothing is known about the dynamics of sperm competition. In this study, we examined the outcome of sperm competition when virgin female guppies mated with two males. Behavioural cues were used to ensure that each male mated once (with female cooperation) and that sperm were successfully inseminated at copulation. Two polymorphic microsatellite loci were used to estimate the proportion of offspring sired by the second male (P2) and the results revealed a bimodal distribution with either first or (more often) second male priority The observed P2 distribution differed from that expected under the 'fair raffle' model of sperm competition. Random sperm mixing is therefore unlikely to account for the observed variance in P2 in this study. A further aim of our study was to identify predictors of male reproductive success. Using logistic linear modelling, we found that the best predictors of paternity were time to remating and the difference in courtship display rate between first and second males. Males that mated quickly and performed relatively high numbers of sigmoid displays obtained greater parentage than their slower and less vigorous counterparts. Since females are attracted to high-displaying males, our results suggest that female choice may facilitate sperm competition and/or sperm choice in guppies.

The African Caribbean eye survey: risk factors for glaucoma in a sample of African Caribbean people living in London

The purpose of the study was to estimate the prevalence of and risk factors for chronic glaucoma in a sample of African Caribbean people over 35 years of age living in the London Borough of Haringey. A cross-sectional voluntary sample of persons were subjected to detailed ophthalmic assessment including automated tangent screen suprathreshold visual field testing, applanation tonometry and stereoscopic disc evaluation in 50 community-based survey clinics over an 8 month period. Cases and suspects were referred to Moorfields Eye Hospital for more detailed assessment and confirmation of the diagnosis. Of 873 eligible persons examined (out of a total of 1022), 32 definite cases of glaucoma were identified, a prevalence of 3.9 percent; 42 percent of these had been previously diagnosed. Approximately 10 percent of the sample required further assessment and follow-up when ocular hypertensives and glaucoma suspects were included. An age-standardised comparison with the findings of the Roscommon survey revealed a relative risk for glaucoma for Haringey blacks compared with Irish whites of 3.7. Significant risk factors for glaucoma included age, African birthplace and darker skin colour. Neither diabetes nor hypertension reached significance. Despite the lack of a population base, this study provides strong evidence that the 4 times greater risk of glaucoma estimated for American blacks compared with whites applies equally to the United Kingdom population. Community-based facilities are required to raise awareness of the risk among this ethnic minority in this country and case-finding resources should be provided to meet local needs (Summary)

Early language and communicative abilities of children with periventricular leukomalacia.

Ten 2-year-old children with periventricular leukomalacia (PVL), a brain injury associated with prematurity, were evaluated using language samples. Five children scored less than 80 on cognitive testing (delayed). Five children with this disorder and normal cognitive scores were assessed at two ages, matched with the delayed group on CA and developmental level. The delayed group produced significantly fewer lexical tokens and spontaneous verbal utterances than did the CA-matched group. No significant differences were observed between the delayed group and either comparison group on other measures of lexicon, grammar, or communication. The data demonstrate a relation between cognitive abilities and measures of verbal productivity in children with PVL.

Physicians play key role in information systems planning.

Healthcare executives struggle with schemes to encourage their institution's affiliated physicians to "buy" into the information systems master plan. By involving physicians initially as members of an MIS Executive Committee, one tertiary-care medical center in Pennsylvania promoted not only "buy-in," but stimulated physician curiosity, active use and even demands about future directions of IS at the medical center.

Effect of high-dose vitamin D supplementation on radiographically detectable bone disease of very low birth weight infants.

To test the hypothesis that high-dose vitamin D2 supplementation would result in a lower incidence of radiographically detectable bone disease, we randomly assigned 40 very low birth weight infants to a control group who received vitamin D2 in a dosage of 400 IU/day and 41 to an experimental group who received a dosage of 2000 IU/day. After 6 weeks, radiographs from all infants were scored blindly for degree of radiographic bone disease, and serum osteocalcin and 25-hydroxyvitamin D levels were measured. Mean vitamin D intake was 360 +/- 141 (SD) IU/day in the control group and 2170 +/- 144 (SD) IU/day in the experimental group. Median 6-week serum 25-hydroxyvitamin D levels were 24 ng/ml (range 3 to 60 ng/ml) in the control group and 68 ng/ml (range 9 to 150 ng/ml) in the experimental group (p less than 0.001). Overall, 20% of the infants had evidence of moderate radiographic bone disease and only 2% were severely affected. The radiographic bone score (median = 2.5) and serum osteocalcin concentration (mean = 21.7 +/- 8.7 ng/ml) in the control subjects did not differ significantly from those in the experimental group (median bone score = 2.0; mean osteocalcin level = 24.1 +/- 7.9 ng/ml). Although there may be a subset of very low birth weight infants who would benefit from high doses of vitamin D, we conclude that no generalized clinical improvement can be attributed to this regimen alone.

Effect of timing of cerebral ultrasonography on the prediction of later neurodevelopmental outcome in high-risk preterm infants.

To determine the predictive value of cranial ultrasonographic examination in high-risk preterm infants at different postnatal ages, we scanned 110 infants less than or equal to 32 weeks gestational age at 1, 2, 3, and 6 weeks postnatal ages and at 40 weeks postconceptional age (PCA). Cranial abnormalities detected by ultrasonography at each postnatal age of examination were classified as minor (periventricular superolateral echogenicity with or without intraventricular hemorrhage, grades 1 to 3) or major (cystic periventricular leukomalacia with or without intraventricular hemorrhage, grade 4) and correlated with neurodevelopmental outcome determined by 1 year of age. Major abnormalities detected by ultrasonography were present in four infants at 1 week, four at 2 weeks, eight at 3 weeks, and 11 infants at 6 weeks and 40 weeks PCA, respectively. Nineteen infants (17%) had moderate to severe functional handicaps defined as cerebral palsy, cognitive or visual deficit, or deafness. The positive and negative predictive values of ultrasound examinations, with regard to later neurodevelopmental outcome, improved with increasing postnatal age at examination and was best at 40 weeks PCA. Negative results of ultrasound study at 40 weeks PCA most correctly predicted satisfactory outcome. Although only 58% of moderately to severely handicapped infants were correctly identified by ultrasound examination at 40 weeks PCA, all infants with major ultrasonographic abnormalities at 40 weeks PCA had moderate or severe handicap. Our data demonstrate that the timing of cerebral ultrasonography is important in the prediction of later neurodevelopmental outcome in high-risk preterm infants.