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Background: High hand hygiene (HH) workload is a commonly cited barrier to optimal HH performance. The objective of this study was to assess trends of HH workload as defined by HH opportunities (HHO) and performance rates over different timescales using automated HH monitoring system data., Methods: This multiyear retrospective observational study was conducted in 58 inpatient units located in 10 North American hospitals. HHO and HH rates were analyzed by time series mixed effects general additive model., Results: Median HH rates peaked at 50.0 between 6 and 7 AM with a trough of 38.2 at 5 PM. HHO over hours in a day were the highest at 184 per hospital unit per hour at 10 AM with a trough of 49.0 between 2 and 3 AM. Median rates for day and night shifts were 40.8 and 45.5, respectively (P = .078). Weekend day shift had the lowest median rate (39.4) compared with any other 12-hour shift (P < .1018). The median rates and HHO varied little across days in a week and months., Conclusions: HH workload and performance rates were negatively correlated and changed drastically over hours in a day. Hospitals should consider HH workload in the development and timely delivery of improvement interventions., (Copyright © 2024 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.)

Parasitoid wasps are exceptionally diverse and use specialized adaptations capable of manipulating the physiology and behaviour of host organisms 1 . In more than two centuries since the first records of Drosophila-parasitizing wasps, nearly 200 described and provisional parasitoid species of drosophilids have been identified 2 . These include endoparasitoids and ectoparasitoids, as well as species attacking larval and pupal hosts 3 . Despite a deep history of research attention and remarkable biodiversity, a wasp species that attacks and develops inside the adult stage of a fly host has not been described previously. Here we report the discovery of a wasp species that infects the adult stage of fruit flies in the genus Drosophila, including one of the most deeply studied model organisms in biology, Drosophila melanogaster. Notably, this wasp can be easily collected from backyard fly baits and has a broad geographic distribution throughout the eastern USA. We document its life history and unique host interactions, including egg-laying into and larval emergence from adult flies, and provide protocols to raise wasps from wild-caught host flies. Our results emphasize the need for ongoing research investment in insect biodiversity and systematics. As parasitoid research continues to uncover unusual biology and supports fundamental mechanistic insights into immunity 4 , metabolism 5 , ecology 6 , evolution 7-9 and behaviour 10-12 , we anticipate that this wasp's association with the laboratory model organism, D. melanogaster, will provide new research opportunities across the life sciences., (© 2024. The Author(s).)

Discordance between the genetic sex and phenotype seen on ultrasound can identify disorders of sexual development (DSD) that previously escaped detection until puberty. We describe a 46, XY disorder of sexual differentiation caused by a rare mutation in the SF1 gene (OMIM]184757, ( NR5A1 ). The mutation ( NR5A1 )-c.205C > G (p. Arg69Gly) was discovered after a phenotype-genotype discrepancy was encountered during prenatal care. The baby with 46, XY DSD has female external genitalia but evidence of Y chromosome-related regression of Müllerian structures and the absence of palpable gonads. We discussed the literature on phenotype-genotype discrepancy and the importance of care coordination between the antenatal and postnatal teams to ensure a timely diagnosis of DSD., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2024 Madeline Snipes et al.)

Vertically transmitted (VT) microbial symbionts play a vital role in the evolution of their insect hosts. A longstanding question in symbiont research is what genes help promote long-term stability of vertically transmitted lifestyles. Symbiont success in insect hosts is due in part to expression of beneficial or manipulative phenotypes that favor symbiont persistence in host populations. In Spiroplasma , these phenotypes have been linked to toxin and virulence domains among a few related strains. However, these domains also appear frequently in phylogenetically distant Spiroplasma, and little is known about their distribution across the Spiroplasma genus. In this study, we present the complete genome sequence of the Spiroplasma symbiont of Drosophila atripex , a non-manipulating member of the Ixodetis clade of Spiroplasma , for which genomic data are still limited. We perform a genus-wide comparative analysis of toxin domains implicated in defensive and reproductive phenotypes. From 12 VT and 31 non-VT Spiroplasma genomes, ribosome-inactivating proteins (RIPs), OTU-like cysteine proteases (OTUs), ankyrins, and ETX/MTX2 domains show high propensity for VT Spiroplasma compared to non-VT Spiroplasma . Specifically, OTU and ankyrin domains can be found only in VT- Spiroplasma , and RIP domains are found in all VT Spiroplasma and three non-VT Spiroplasma . These domains are frequently associated with Spiroplasma plasmids, suggesting a possible mechanism for dispersal and maintenance among heritable strains. Searching insect genome assemblies available on public databases uncovered uncharacterized Spiroplasma genomes from which we identified several spaid -like genes encoding RIP, OTU, and ankyrin domains, suggesting functional interactions among those domain types. Our results suggest a conserved core of symbiont domains play an important role in the evolution and persistence of VT Spiroplasma in insects., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Moore and Ballinger.)

Objective: To determine how engagement of the hospital and/or vendor with performance improvement strategies combined with an automated hand hygiene monitoring system (AHHMS) influence hand hygiene (HH) performance rates., Design: Prospective, before-and-after, controlled observational study., Setting: The study was conducted in 58 adult and pediatric inpatient units located in 10 hospitals., Methods: HH performance rates were estimated using an AHHMS. Rates were expressed as the number of soap and alcohol-based hand rub portions dispensed divided by the number of room entries and exits. Each hospital self-assigned to one of the following intervention groups: AHHMS alone (control group), AHHMS plus clinician-based vendor support (vendor-only group), AHHMS plus hospital-led unit-based initiatives (hospital-only group), or AHHMS plus clinician-based vendor support and hospital-led unit-based initiatives (vendor-plus-hospital group). Each hospital unit produced 1–2 months of baseline HH performance data immediately after AHHMS installation before implementing initiatives., Results: Hospital units in the vendor-plus-hospital group had a statistically significant increase of at least 46% in HH performance compared with units in the other 3 groups ( P ≤ .006). Units in the hospital only group achieved a 1.3% increase in HH performance compared with units that had AHHMS alone ( P = .950). Units with AHHMS plus other initiatives each had a larger change in HH performance rates over their baseline than those in the AHHMS-alone group ( P < 0.001)., Conclusions: AHHMS combined with clinician-based vendor support and hospital-led unit-based initiatives resulted in the greatest improvements in HH performance. These results illustrate the value of a collaborative partnership between the hospital and the AHHMS vendor.

Spinal cord stimulation enhanced restoration of motor function following spinal cord injury (SCI) in unblinded studies. To determine whether training combined with transcutaneous electrical spinal cord stimulation (tSCS), with or without systemic serotonergic treatment with buspirone (busp), could improve hand function in individuals with severe hand paralysis following SCI, we assessed ten subjects in a double-blind, sham-controlled, crossover study. All treatments-busp, tSCS, and the busp plus tSCS-reduced muscle tone and spasm frequency. Buspirone did not have any discernible impact on grip force or manual dexterity when administered alone or in combination with tSCS. In contrast, grip force, sinusoidal force generation and grip-release rate improved significantly after 6 weeks of tSCS in 5 out of 10 subjects who had residual grip force within the range of 0.1-1.5 N at the baseline evaluation. Improved hand function was sustained in subjects with residual grip force 2-5 months after the tSCS and buspirone treatment. We conclude that tSCS combined with training improves hand strength and manual dexterity in subjects with SCI who have residual grip strength greater than 0.1 N. Buspirone did not significantly improve the hand function nor add to the effect of stimulation., (© 2022. The Author(s).)

Cytoplasmic incompatibility (CI) is a form of reproductive manipulation caused by maternally inherited endosymbionts infecting arthropods, like Wolbachia , whereby matings between infected males and uninfected females produce few or no offspring. We report the discovery of a new CI symbiont, a strain of Spiroplasma  causing CI in the parasitoid wasp Lariophagus distinguendus . Its extracellular occurrence enabled us to establish CI in uninfected adult insects by transferring Spiroplasma -infected hemolymph. We sequenced the CI- Spiroplasma genome and did not find any homologues of any of the cif genes discovered to cause CI in Wolbachia , suggesting independent evolution of CI. Instead, the genome contains other potential CI-causing candidate genes, such as homologues of high-mobility group (HMG) box proteins that are crucial in eukaryotic development but rare in bacterial genomes. Spiroplasma 's extracellular nature and broad host range encompassing medically and agriculturally important arthropods make it a promising tool to study CI and its applications., Competing Interests: The authors declare no competing interests., (© 2022 The Author(s).)

Objective: To understand the specific ways in which champions lead efforts to obtain and sustain buy-in for immediate postpartum long-acting reversible contraception (LARC) programs., Methods: We conducted a qualitative study with 60 semistructured interviews at 3 teaching hospitals in Texas with physicians, nurses, administrators and other staff who participated in the implementation of immediate postpartum LARC. Physicians self-identified as champions and identified other champion physicians and administrators. Two researchers analyzed and coded interview transcripts for content and themes., Results: We found that champions draw on institutional knowledge and relationships to build awareness and support for immediate postpartum LARC implementation. To obtain buy-in, champions needed to demonstrate financial sustainability, engage key stakeholders from multiple departments, and obtain nurse buy-in. Champions also created buy-in by communicating goals for the service that focused on expanding reproductive autonomy, improving maternal health, and improving access to postpartum contraception. Some staff, especially nurses, identified reasons for the program that run counter to reproductive justice principles: reducing birth rates, poverty, and/or unplanned pregnancy among young women and high-parity women. Respondents at 2 hospitals noted that not all women had equitable access to immediate postpartum LARC., Conclusion: Physician and non-physician champions must secure long-term support across multiple hospital departments to successfully implement an immediate postpartum LARC program. For programs to equitably serve all women in need of postpartum contraceptive care, champions and other program leaders need to implement strategies to address access issues. They should also explicitly focus on reproductive justice principles during program introduction and training., Implications: Successfully implementing immediate postpartum long-acting reversible contraception programs requires champions with institutional networking connections, administrative and nursing support, and clearly communicated goals. Champions need to address access issues and focus on reproductive justice principles during program introduction and training to equitably serve all women in need of postpartum contraceptive care., (Copyright © 2021. Published by Elsevier Inc.)

Inherited mutualists, parasites, and commensals occupy one of the most intimate ecological niches available to invertebrate-associated microbes. How this transmission environment influences microbial evolution is increasingly understood for inherited bacterial symbionts, but in viruses, research on the prevalence of vertical transmission and its effects on viral lineages is still maturing. The evolutionary stability of this strategy remains difficult to assess, although phylogenetic evidence of frequent host shifts and selective sweeps have been interpreted as strategies favoring parasite persistence. In this study, we describe and investigate a natural insect system in which species-wide sweeps have been restricted by the isolation of host populations. Previous work identified evidence of pronounced mitochondrial genetic structure among North American populations of the phantom midge, Chaoborus americanus . Here we take advantage of the geographical isolation in this species to investigate the diversity and persistence of its inherited virome. We identify eight novel RNA viruses from six families and use small RNA sequencing in reproductive tissues to provide evidence of vertical transmission. We report region-specific virus strains that mirror the continental phylogeography of the host, demonstrating that members of the inherited virome have independently persisted in parallel host lineages since they last shared a common ancestor in the Mid-Pleistocene. We find that the small interfering RNA pathway, a frontline of antiviral defense in insects, targets members of this inherited virome. Finally, our results suggest that the Piwi-mediated RNA silencing pathway is unlikely to function as a general antiviral defense in Chaoborus , in contrast to its role in some mosquitoes. However, we also report that this pathway generates abundant piRNAs from endogenous viral elements closely related to actively infecting inherited viruses, potentially helping to explain idiosyncratic patterns of virus-specific Piwi targeting in this insect., (© The Author(s) 2022. Published by Oxford University Press.)

Background: Achieving high levels of hand hygiene compliance of health care personnel has been an ongoing challenge. The objective of this study was to examine the impact of the COVID-19 pandemic on hand hygiene performance (HHP) rates in acute care hospitals., Methods: HHP rates were estimated using an automated hand hygiene monitoring system installed in 74 adult inpatient units in 7 hospitals and 10 pediatric inpatient units in 2 children's hospitals. A segmented regression model was used to estimate the trajectory of HHP rates in the 10 weeks leading up to a COVID-19-related milestone event (eg, school closures) and for 10 weeks after., Results: Three effects emerged, all of which were significant at P < .01. Average HHP rates increased from 46% to 56% in the months preceding pandemic-related school closures. This was followed by a 6% upward shift at the time school closures occurred. HHP rates remained over 60% for 4 weeks before declining to 54% at the end of the study period., Conclusions: Data from an automated hand hygiene monitoring system indicated that HHP shifted in multiple directions during the early stages of the pandemic. We discuss possible reasons why HHP first increased as the pandemic began and then decreased as it progressed., (Copyright © 2020 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.)

Objective: To determine the effect of using a technician-driven medication therapy management (MTM) program on quality performance measures for a community pharmacy chain., Methods: A technician-driven MTM program was incorporated in 35 stores of a regional supermarket pharmacy chain. The overall chain percentage score for the 4 quality measures used in Medicare Part D Star Ratings-proportion of days covered (PDC) for cholesterol, diabetes, renin-angiotensin system antagonists (RASA), and high-risk medication use-was compared pre- and postimplementation of the technician-driven MTM program. Data were collected from Electronic Quality Improvement Platform for Plans and Pharmacies (EQuIPP) platform and t tests were used to analyze 1 year pre- and postimplementation., Results: The PDC for RASA, high cholesterol medications, and diabetes medications significantly increased pre- to postimplementation for each drug class ( P < .001, P = .011, P = .001, respectively). The combined overall mean PDC score for RASA, cholesterol medications, and diabetes medication classes significantly increased by 5.6% from 2015 to 2016 (74.2% vs 79.8%, P < .001); there was also a nonsignificant decrease in high-risk medication use for the entire chain., Conclusions: This technician-driven MTM program can positively affect pharmacy quality performance and potentially improve patient outcomes.

Microbial partners play important roles in the biology and ecology of animals. In insects, maternally transmitted symbionts are especially common and can have host effects ranging from reproductive manipulation to nutrient provisioning and defense against natural enemies. In this study, we report a genus-wide association of Myrmica ants with the inherited bacterial symbiont Spiroplasma We screen Myrmica ants collected from the wild, including the invasive European fire ant, Myrmica rubra , and find an extraordinarily high prevalence of this symbiont-8 of 9 species, 42 of 43 colonies, and 250 of 276 individual workers harbored Spiroplasma -only one host species was uninfected. In our screens, each host species carried a distinct Spiroplasma strain, and none were infected with more than one strain. All symbionts belong to the citri clade, allied most closely with pathogenic strains of Spiroplasma infecting corn crops and honeybees, and there is strong evidence of host-symbiont persistence across evolutionary time scales. Genome sequencing of two Spiroplasma symbionts revealed candidate genes that may play a part in the symbiosis, a nutrient transporter absent from other Spiroplasma strains, and a ribosome-inactivating protein previously implicated in parasite defense. These results together suggest long-term, likely mutualistic, relationships atypical of Spiroplasma -insect associations with potential significance for broad ecological interactions with Myrmica IMPORTANCE Animal-associated microbial symbionts can dramatically affect the biology of their hosts. The identification and characterization of these intimate partnerships remain an essential component of describing and predicting species interactions, especially for invasive host species. Ants perform crucial ecological functions as ecosystem engineers, scavengers, and predators, and ants in the genus Myrmica can be aggressive resource competitors and reach high densities in their native and invaded habitats. In this study, a novel symbiosis is identified between Myrmica ants and the facultative bacterial symbiont Spiroplasma Broad host distribution, high frequencies of infection, and host-symbiont codivergence over evolutionary time scales, an uncommon feature of Spiroplasma associations, suggest an important likely mutualistic interaction. Genome sequencing identified highly divergent gene candidates that may contribute to Spiroplasma 's role as a possible defensive or nutritional partner in Myrmica ., (Copyright © 2018 American Society for Microbiology.)

Candida spp . are opportunistic unicellular fungi, known to cause oral, vaginal, lung and occasionally systemic infections. Characteristically, they colonize the oral cavity, the mucosal surfaces of the cheek, palate, and tongue. Usually harmless, oral Candidas may become pathogenic under immunosuppressive conditions, dentures presence, or salivary flow impairment. Accurate species identification is important because C. dubliniensis can rapidly develop fluconazole resistance. We report C. dubliniensis in an immunocompetent patient with a metal lingual frenulum piercing.

In the mammalian genome, DNA methylation is an epigenetic mechanism involving the transfer of a methyl group onto the C5 position of the cytosine to form 5-methylcytosine. DNA methylation regulates gene expression by recruiting proteins involved in gene repression or by inhibiting the binding of transcription factor(s) to DNA. During development, the pattern of DNA methylation in the genome changes as a result of a dynamic process involving both de novo DNA methylation and demethylation. As a consequence, differentiated cells develop a stable and unique DNA methylation pattern that regulates tissue-specific gene transcription. In this chapter, we will review the process of DNA methylation and demethylation in the nervous system. We will describe the DNA (de)methylation machinery and its association with other epigenetic mechanisms such as histone modifications and noncoding RNAs. Intriguingly, postmitotic neurons still express DNA methyltransferases and components involved in DNA demethylation. Moreover, neuronal activity can modulate their pattern of DNA methylation in response to physiological and environmental stimuli. The precise regulation of DNA methylation is essential for normal cognitive function. Indeed, when DNA methylation is altered as a result of developmental mutations or environmental risk factors, such as drug exposure and neural injury, mental impairment is a common side effect. The investigation into DNA methylation continues to show a rich and complex picture about epigenetic gene regulation in the central nervous system and provides possible therapeutic targets for the treatment of neuropsychiatric disorders.

JC virus (JCV) is the aetiological agent of the demyelinating disease progressive multifocal leukoencephalopathy, an AIDS defining illness and serious complication of mAb therapies. Initial infection probably occurs in childhood. In the working model of dissemination, virus persists in the kidney and lymphoid tissues until immune suppression/modulation causes reactivation and trafficking to the brain where JCV replicates in oligodendrocytes. JCV infection is regulated through binding of host factors such as Spi-B to, and sequence variation in the non-coding control region (NCCR). Although NCCR sequences differ between sites of persistence and pathogenesis, evidence suggests that the virus that initiates infection in the brain disseminates via B-cells derived from latently infected haematopoietic precursors in the bone marrow. Spi-B binds adjacent to TATA boxes in the promoter/enhancer of the PML-associated JCV Mad-1 and Mad-4 viruses but not the non-pathogenic, kidney-associated archetype. The Spi-B-binding site of Mad-1/Mad-4 differs from that of archetype by a single nucleotide, AAAAGGGAAGGGA to AAAAGGGAAGGTA. Point mutation of the Mad-1 Spi-B site reduced early viral protein large T-antigen expression by up to fourfold. Strikingly, the reverse mutation in the archetype NCCR increased large T-antigen expression by 10-fold. Interestingly, Spi-B protein binds the NCCR sequence flanking the viral promoter/enhancer, but these sites are not essential for early viral gene expression. The effect of mutating Spi-B-binding sites within the JCV promoter/enhancer on early viral gene expression strongly suggests a role for Spi-B binding to the viral promoter/enhancer in the activation of early viral gene expression.

Aberrant hyaluronan production has been implicated in many types of tumor. In this context, hyaluronidase has been explored as a viable therapeutic approach to reduce tumoral hyaluronan. However, elevated levels of hyaluronan in tumors are often associated with high expression levels of cellular hyaluronidases, which consequently produce various sizes of saturated hyaluronan fragments with divergent pro-tumoral activities. The current study shows that different hyaluronan metabolisms of mammalian and microbial hyaluronidases could elicit distinct alterations in cancer cell behavior. Unlike saturated hyaluronan metabolites, unsaturated hyaluronan oligosaccharides produced by bacteriophage hyaluronidase, HylP, had no biological effect on growth of breast carcinoma cells. More importantly, HylP's metabolic process of hyaluronan into non-detrimental oligosaccharides significantly decreased breast cancer cell proliferation, migration and invasion by disrupting Erk1/2 activation and RhoA expression. Our results suggest that it may be possible to exploit HylP's unique enzymatic activity in suppressing hyaluronan-mediated tumor growth and progression., (Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.)

Background: Although anti-angiogenic therapy is a promising new line of therapy for prostate cancer, we recently reported that stable expression of endostatin arrested the progression of prostate cancer to poorly differentiated state and distant metastasis in TRAMP mice. However, the same therapy failed to provide any benefit when given either during or after the onset of metastatic switch. The present study determined the possible mechanisms behind the selective advantage of endostatin therapy in early-stage disease., Methods: Angiogenesis-related gene expression analysis was performed to identify target genes and molecular pathways involved in the therapy effects. Based on the results from in vivo studies, and recapitulation of the in vivo data in vitro using tumorigenic and non-tumorigenic human prostate cancer cells that are either androgen-sensitive or androgen-independent, analyses of possible mechanisms of the selective advantage of early treatment were performed using assays for cell proliferation, apoptosis, migration, and cell signaling. The identified mechanisms were further confirmed in vivo., Results: Results indicated that cells with high androgen receptor (AR) expression were more sensitive to endostatin treatment than androgen-independent cells with low or no AR expression. Endostatin was found to significantly downregulate the expression of growth factors, receptor tyrosine kinases, proteases, and AR both in vitro and in vivo only when the cells express high-levels of AR. Cell proliferation was not influenced by endostatin treatment but migration was significantly affected only in androgen-sensitive cells. Targeted downregulation of AR prior to endostatin treatment in androgen-sensitive cells and overexpression of AR in androgen-independent cells indicated that the effect of endostatin via AR downregulation is mediated by a non-genotropic mechanism on Ras and RhoA pathways, and independently of AR on MAPK/ERK pathway., Conclusions: These data indicate that systemically stable endostatin expression delays the onset of metastatic switch by acting on multiple pathways involving AR., ((c) 2009 Wiley-Liss, Inc.)

Purpose: Overexpression of transforming growth factor (TGF)-beta has been implicated in promoting immune suppression, tumor angiogenesis, tumor cell migration, and invasion in many cancers, including carcinoma of the breast. Thus, targeted down-regulation of TGF-beta1 expression in breast cancer in situ and determination of its implications would provide new treatment approaches for disease management., Experimental Design: Small interfering RNA constructs targeting TGF-beta1 were validated and used to develop clonal derivatives of the metastatic breast cancer cell line MDA-MB-435. The cells were used in several in vitro analyses, including migration, invasion, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, apoptosis, and signaling assays. A wound-healing assay was used to determine migration of the cells in culture and a Boyden chamber transwell assay was used for invasion. Further, the clones were used in an in vivo mouse model for the kinetics of tumor growth and gene expression in the primary site and in lungs upon metastasis., Results: Inhibition of TGF-beta1 expression in MDA-MB-435 cells showed a 35% decrease in migration and a 55% decrease in invasion in vitro, with a 50% increase in proliferation and no effect on apoptosis. In vivo analysis indicated a 90% decrease in the number of mice bearing macroscopic lung metastases; however, the primary tumors did not show any difference in the growth kinetics when compared with the parental MDA-MB-435 cells. Analysis of TGF-beta signaling pathways in the clonal derivatives showed a decrease in Smad2 activation and an increase in AKT and extracellular signal-regulated kinase activation. Interestingly, analysis of TGF-beta receptor expression showed a decrease in both receptor I and II expression in TGF-beta1 silenced cells. These results suggest that inhibition of TGF-beta1 ligand may act as a negative feedback loop to disrupt the function of all TGF-beta isoforms., Conclusions: Therapies targeting the TGF-beta signaling pathway may be more effective in late-stage disease to prevent organ metastasis but not primary tumor formation and may be combined with other tumor-targeted therapies normally limited by increased circulating TGF-beta levels.

Bacterial artificial chromosome (BAC) clones from apomicts Pennisetum squamulatum and buffelgrass (Cenchrus ciliaris), isolated with the apospory-specific genomic region (ASGR) marker ugt197, were assembled into contigs that were extended by chromosome walking. Gene-like sequences from contigs were identified by shotgun sequencing and BLAST searches, and used to isolate orthologous rice contigs. Additional gene-like sequences in the apomicts' contigs were identified by bioinformatics using fully sequenced BACs from orthologous rice contigs as templates, as well as by interspecies, whole-contig cross-hybridizations. Hierarchical contig orthology was rapidly assessed by constructing detailed long-range contig molecular maps showing the distribution of gene-like sequences and markers, and searching for microsyntenic patterns of sequence identity and spatial distribution within and across species contigs. We found microsynteny between P. squamulatum and buffelgrass contigs. Importantly, this approach also enabled us to isolate from within the rice (Oryza sativa) genome contig Rice A, which shows the highest microsynteny and is most orthologous to the ugt197-containing C1C buffelgrass contig. Contig Rice A belongs to the rice genome database contig 77 (according to the current September 12, 2003, rice fingerprint contig build) that maps proximal to the chromosome 11 centromere, a feature that interestingly correlates with the mapping of ASGR-linked BACs proximal to the centromere or centromere-like sequences. Thus, relatedness between these two orthologous contigs is supported both by their molecular microstructure and by their centromeric-proximal location. Our discoveries promote the use of a microsynteny-based positional-cloning approach using the rice genome as a template to aid in constructing the ASGR toward the isolation of genes underlying apospory.

A "gene-island" sequencing strategy has been developed that expedites the targeted acquisition of orthologous gene sequences from related species for comparative genome analysis. A 152-kb bacterial artificial chromosome (BAC) clone from sorghum (Sorghum bicolor) encoding phytochrome A (PHYA) was fully sequenced, revealing 16 open reading frames with a gene density similar to many regions of the rice (Oryza sativa) genome. The sequences of genes in the orthologous region of the maize (Zea mays) and rice genomes were obtained using the gene-island sequencing method. BAC clones containing the orthologous maize and rice PHYA genes were identified, sheared, subcloned, and probed with the sorghum PHYA-containing BAC DNA. Sequence analysis revealed that approximately 75% of the cross-hybridizing subclones contained sequences orthologous to those within the sorghum PHYA BAC and less than 25% contained repetitive and/or BAC vector DNA sequences. The complete sequence of four genes, including up to 1 kb of their promoter regions, was identified in the maize PHYA BAC. Nine orthologous gene sequences were identified in the rice PHYA BAC. Sequence comparison of the orthologous sorghum and maize genes aided in the identification of exons and conserved regulatory sequences flanking each open reading frame. Within genomic regions where micro-colinearity of genes is absolutely conserved, gene-island sequencing is a particularly useful tool for comparative analysis of genomes between related species.

SUMMARY Previous research has demonstrated that those who believe that homosexuality is genetically or biologically caused have less negative attitudes towards gays and lesbians than those who believe it is acquired, learned, or chosen. This study, utilizing an undergraduate and graduate Psychology student sample, found significant relationships between attitudes towards lesbians and gay men and beliefs about causes and "treatments" for homosexuality. Level of personal contact with lesbians and gay men and demographic factors also influenced attitudes toward and beliefs about homosexuality. These results suggest that educational attempts to change attitudes towards lesbians and gay men should consider the role played by beliefs about the etiologies of homosexuality.

Objectives: This study sought to determine prevalence of and risk factors for nonfatal recent overdose among street-recruited injection heroin users., Methods: From August 1998 through July 1999, 1427 heroin injectors were recruited from 6 inner-city neighborhoods in the San Francisco Bay Area, Calif, and interviewed. Factors hypothesized to be associated with recent overdose were analyzed with logistic regression., Results: Of the 1427 participants, 684 (48%) had had an overdose, 466 (33%) had experienced 2 or more overdose events, and 182 (13%) had had a recent overdose. In multiple logistic regression, being younger (adjusted odds ratio [OR] for each year of increasing age = 0.95; 95% confidence interval [CI] = 0.94, 0.97), having been arrested 3 or more times in the past year (adjusted OR = 2.50; 95% CI = 1.61, 3.87), drinking 4 or more alcoholic drinks per day (adjusted OR = 2.05; 95% CI = 1.37, 3.05), and having participated in methadone detoxification during the past year (adjusted OR = 1.47; 95% CI = 1.03, 2.09) were independently associated with recent overdose. Being homeless; identifying as gay, lesbian, bisexual, or transgender; having spent 5 or more years in prison or jail; and having engaged in sex work also were associated with recent overdose., Conclusions: Targeted interventions that decrease risk for overdose are urgently needed.

Directly observed therapy (DOT) has become a popular strategy in tuberculosis (TB) control. Limited resources to provide DOT makes selection of patients an issue. Studies were done in San Francisco to analyze the socioeconomic and behavioral characteristics of 1992 TB patients--and of those on DOT--and to determine attitudes of indigent San Franciscans toward TB and adherence to TB control measures. The studies show that (1) African American and Latino patients were more likely to be selected for DOT than Asian/Pacific Islanders and whites, regardless of factors associated with adherence behavior, and (2) DOT, delivered in a respectful manner and combined with incentives, was evaluated positively by those with risk factors.

Theophylline level testing enables the physician to monitor patients on theophylline and maintain benefit vs risk at an optimum ratio. This study consisted of a retrospective chart review of 53 patients who had a total of 103 serum theophylline level determinations (STLDs) over a 12-month period. The study was designed to look at reasons why physicians ordered STLDs and to what extent those results influenced subsequent theophylline prescribing. Findings showed that a large number of STLDs were ordered on asymptomatic patients with no recent dose change or initiation of therapy and no recent hospitalization or emergency room visit. The most common reason for ordering an STLD was presentation with symptoms or signs of asthma and no other recent events. On several occasions when low results were obtained, theophylline dosage was not increased. In some of these cases the patient's clinical presentation may have influenced the decision to maintain the same dosage. Use of erythromycin and smoking status were observed to affect theophylline clearance. Most physicians failed to document time of last theophylline dose, which hindered accurate interpretation of STLDs.

Streptococcus mutans strain GS-5 was grown in a variety of carbon sources in order to achieve different balanced growth rates, ranging from 61 to 383 minutes. The influence of the S. mutans growth rate on mouse phagocyte activity against these bacteria has been evaluated. The percentages of bacteria phagocytized and intracellularly killed by macrophages rose to 60-80% and 85-95% respectively when the doubling time was longer, showing that S. mutans is particularly sensitive to nonspecific immune defence mechanisms when cultured under conditions similar to those of its natural ecosystem.

In Shenandoah National Park, O(3) monitoring data were characterized and attempts were made to relate O(3) concentration levels to visible foliar injury observed for five plant species surveyed. Foliar injury for three species increased with elevation. The 24-h monthly mean O(3) concentrations tended to increase with elevation; however, the number of elevated hourly occurrences did not. Although the frequency of high hourly O(3) concentrations did not consistently increase with elevation, O(3) exposures in the park may have been high enough to provoke an effect that may have been enhanced by vegetation sensitivities that differed as a function of altitude.

Eye movements of two patients with Duane's retraction syndrome (type 1) were recorded using high-resolution infrared oculography. Slowed hypometric abduction was found. The dynamics of adducting saccades in the affected hypometric eye were normal, suggesting an absence of functional cocontraction of the medial and lateral rectus muscles. Therefore, narrowing of the palpebral tissue on adduction most likely reflects a reorganization of the central ocular motor pathways.

'Wando' pea is susceptible to Ditylenchus dipsaci from Raleigh, N. C. (RNC) but resistant to the same species from Waynesville, N. C. (WNC). Homogenates of RNC and WNC were analyzed for pectolytic and cellulolytic enzyme activity; both had high C(x) activity with WNC two to three times more active than RNC. Polymethylglacturonase activity was three to five times higher in RNC, but polygalacturonase was up to 100 times higher in WNC. Polygalacturonate-trans-eliminase was not detected although a Ca(++)-stimulated pectin methyl-trans-eliminase was present. Enzyme analyses of healthy and infected pea tissue showed only slight enzyme activity unrelated to that in nematode homogenates. No correlation between enzyme activity and the differing pathogenicities could be detected.

Moore, L. Daneo (University of Pennsylvania, Philadelphia) and D. J. O'Kane. Kinases of Streptococcus faecalis. I. Enzyme localization. J. Bacteriol. 86:766-772. 1963.-Cell-free extracts of Streptococcus faecalis 10C1 were found to contain a membrane-associated, particulate hexokinase with activity toward glucose, fructose, mannose, and, to a lesser extent, galactose, gluconate, and mannitol. In addition, the cell cytoplasm contained kinases specific for fructose, mannose, and galactose. No specific kinases were found for gluconate or mannitol, although in the presence of mannitol adenosine triphosphate was utilized by a cytoplasmic fraction. The possibility is discussed that the main pathways of gluconate and mannitol metabolism are oxidation followed by phosphorylation.

In cell-free extracts derived from Streptococcus faecalis, protein synthesis directed by endogenous messenger RNA increases as the culture ages. The increased activity is accompanied by an increase in the percentage of membranebound ribosomes and by a decrease in ribosomal monomers and subunits. These changes progress against a background of structural and compositional modifications in the membrane. Membrane modifications possibly related to endogenously directed protein synthesis in cell-free extracts include: (i) decreased specific activity of a membrane-associated polynucleotide phosphorylase capable of polysome degradation, and (ii) increased concentrations of certain phospholipids.