Search

The budding of the urogenital sinus epithelium into the surrounding mesenchyme signals the onset of prostate morphogenesis. The epithelial and mesenchymal factors that regulate ductal budding and the ensuing process of ductal growth and branching are not fully known. We provide evidence that bone morphogenetic protein 4 (BMP4) is a mesenchymal factor that regulates ductal morphogenesis. The Bmp4 gene was most highly expressed in the male urogenital sinus from embryonic day 14 through birth, a period marked by formation of main prostatic ducts and initiation of ductal branching. From an initial wide distribution throughout the prostatic anlage of the urogenital sinus, Bmp4 expression became progressively restricted to the mesenchyme immediately surrounding the nascent prostatic ducts and branches. Exogenous BMP4 inhibited epithelial cell proliferation and exhibited a dose-dependent inhibition of ductal budding in urogenital sinus tissues cultured in vitro. Adult Bmp4 haploinsufficient mice exhibited an increased number of duct tips in both the ventral prostate and coagulating gland. Taken together, our data indicate that BMP4 is a urogenital sinus mesenchymal factor that restricts prostate ductal budding and branching morphogenesis. Key Words: bone morphogenetic protein 4; prostate development; branching morphogenesis.

An automated microsphere-based flow cytometric assay (FlowMetrix™ system) was compared with a conventional enzyme-linked immunosorbent assay (ELISA) for quantifying Ig classes in serum and stool samples. The reproducibility of the process of coupling capture antibodies to microspheres was tested. The use of independently coupled microspheres did not increase the variation of assay results relative to using the same bead set in repeated assays. However, it is necessary to ensure quality control of the coupling process since slight variations in the coupling procedures can profoundly affect the density of capture reagents coupled to the microspheres and consequently adversely affect assay precision. Although the ELISA was more sensitive and did not have the problems with instrument performance encountered with the FlowMetrix™ assay, the latter was more reproducible, had a greater dynamic range of measurement, and took considerably less preparation time than the ELISA. Greater reproducibility is especially important for measurement of fecal Ig, which is typically highly variable. Thus, in addition to its multi-analyte capability, the FlowMetrix™ assay system has definite advantages over a conventional ELISA. Mechanical problems such as microspheres settling to the bottom of wells during analysis by an automated plate reader will likely be overcome, and sensitivity improved as this technology develops. [Copyright &y& Elsevier]

Interest in understanding the environmental distribution of the alkane monooxygenase (AlkB) enzyme led to the identification of over 100 distinct alkane monooxygenase (AlkB) enzymes containing a covalently bound, or fused, rubredoxin. The rubredoxin-fused AlkB from Dietzia cinnamea was cloned as a full-length protein and as a truncated protein with the rubredoxin domain deleted. A point mutation (V91W) was introduced into the full-length protein, with the goal of assessing how steric bulk in the putative substrate channel might affect selectivity. Based on activity studies with alkane and alkene substrates, the rubredoxin-fused AlkB oxidizes a similar range of alkane substrates relative to its rubredoxin domain-deletion counterpart. Oxidation of terminal alkenes generated both an epoxide and a terminal aldehyde. The products of V91W-mutant-catalyzed oxidation of alkenes had a higher aldehyde-to-epoxide ratio than the products formed in the presence of the wild type protein. These results are consistent with this mutation causing a structural change impacting substrate positioning. [Display omitted] • Alkane monooxygenases with fused rubredoxin widely distributed in nature • Rubredoxin deletion mutant has similar substrate range to wild type • Single point mutation leads to an enzyme with substantial anti-Markovnikov activity on alkenes [ABSTRACT FROM AUTHOR]

Underage drinking is an important public health issue for American Indian and Alaska Native (AI/AN) adolescents, as it is for U. S. teens of all ethnicities. One of the demonstrated risk factors for the development of alcohol use disorders in AI/AN is early age of initiation of drinking. To address this issue a randomized trial to assess the efficacy of Motivational Interviewing (MI) compared to Psycho-Education (PE) to reduce and prevent underage drinking in AI/AN youth was developed and implemented. Sixty-nine youth received MI or PE and 87% were assessed at follow-up. For teens who were already drinking, participating in the intervention (MI or PE) was associated, at follow-up, with lower quantity×frequency (q×f) of drinking (p=0.011), fewer maximum drinks per drinking occasion (p=0.004), and fewer problem behaviors (p=0.009). The MI intervention resulted in male drinkers reporting a lower q×f of drinking (p=0.048) and female drinkers reporting less depression (p=0.011). In teens who had not started drinking prior to the intervention, 17% had initiated drinking at follow-up. As a group they reported increased quantity×frequency of drinking (p=0.008) and maximum drinks (p=0.047), but no change in problem behaviors. These results suggest that intervening against underage drinking using either MI or PE in AI/AN youth can result in reduced drinking, prevention of initiation of drinking, and other positive behavioral outcomes. Brief interventions that enhance motivation to change as well as Psycho-Education may provide a successful approach to reducing the potential morbidity of underage drinking in this high-risk group. [ABSTRACT FROM AUTHOR]

Background: To amplify earlier studies of unintended consequences of public policies, this article illustrates both negative and positive unanticipated consequences of smoke-free workplace policies in California bars for women of low SES. Methods: The article relies on thematic analysis in 2008 of qualitative data gathered between 2001 and 2007 from three mixed-method studies of tobacco use in and around bars where indoor smoking is prohibited. Results: Unanticipated consequences primarily occurred when bars did comply with the law and smokers went outside the bar to smoke, particularly when smokers stood on the street outside the bar. Key negative consequences for women who smoked outside of bars included threats to their physical safety and their public image. For women living near bars, increased smoking on the street may have increased their exposure to secondhand smoke and disruptive noise. For some women, however, unanticipated negative consequences were identified with noncompliant bars. Smokers were conjectured to congregate in the smaller number of bars where smoking was still allowed, resulting in increased exposure to secondhand smoke for low-SES women working in these bars. A common positive unintended consequence of the tobacco control ordinance was increased social circulation and solidarity, as smokers gathered outside bars to smoke. Conclusions: Smoke-free workplace laws in bars can have both negative and positive consequences for workers and smokers, and low-income women in particular. [Copyright &y& Elsevier]