Chemical components and their locations in the Verticillium fungicola cell wall

The chemical structure of cell walls and fractions of Verticillium fungicola, a pathogen of Agaricus bisporus, as well as their corresponding ultrastructures were studied. There are at least three chemically distinct types of carbohydrate polymers: one yielding mannose with lower amounts of galactose and glucose (glucogalactomannan), another one composed mainly of glucose (glucan), and a third one containing only N-acetylglucosamine (chitin). Attempts were made to locate these materials in situ by comparing electron micrographs of shadowed and sectioned cell walls, and also by indirect immunofluorescence. It was shown that none of these polymers constituted a completely physically distinct layer, but there seem to be different solubility properties in the outer, inner, and intermediate layers. It was also shown that fibrillar material (chitin) embedded in cementing glucan constituted the residual inner fraction of the original wall material. Indirect immunofluorescence showed the location of a significant amount of glucogalactomannan on the surface of the walls in which rodlet structures were visualized by electron microscopy.     

alpha-Crystallin facilitates the reactivation of hydrogen peroxide-inactivated rhodanese

It was previously shown that rhodanese, inactivated with hydrogen peroxide, could only be reactivated in the presence of a reductant or the substrate thiosulfate if these reagents were added soon after inactivation and if the oxidant was removed. Here, we report on the facilitated reactivation (75%) of hydrogen peroxide-inactivated rhodanese by the chaperone alpha-crystallin. Reactivation by the chaperone still required a reductant and thiosulfate. Without alpha-crystallin, but in the presence of the reductant and thiosulfate, the inactivated enzyme regained about 39% of its original activity. The alpha-crystallin-assisted reactivation of hydrogen peroxide-inactivated rhodanese was independent of ATP. Further, we found, that alpha-crystallin interacted transiently, but could not form a stable complex with hydrogen peroxide-inactivated rhodanese. Unlike in prior studies that involved denaturation of rhodanese through chemical or thermal means, we have clearly shown that alpha-crystallin can function as a molecular chaperone in the reactivation of an oxidatively inactivated protein.     

Influences of blood sampling procedures on basal hypothalamic-pituitary-adrenal hormone levels and leukocyte values in rhesus macaques (Macaca mulatta)

Abstract: The influences of housing location and temporal factors on the results of blood sampling were examined to determine their contributions to levels of hypothalamic-pituitary-adrenal hormones, as well as leukocyte subset counts from peripheral blood in rhesus monkeys. Differences in housing location and the amount of room disturbance associated with blood sampling have a significant impact on cell counts, but not on ACTH or Cortisol levels.     

Sustainability of Evidence-Based Practices for HIV Prevention among Female Sex Workers in Mexico

Objective

This study examined service provider perceptions of requirements for successful sustainment of an efficacious intervention for preventing HIV/AIDS and STIs in female sex workers (FSWs) in Mexico.

Methods

Semi-structured interviews were conducted with 77 leaders and counselors from 12 community-based reproductive health clinics located throughout Mexico participating in a large hybrid effectiveness-implementation randomized controlled trial to scale-up the use of Mujer Segura, a psychoeducational intervention designed to promote condom use and enhance safer sex negotiation skills among FSWs.

Results

Five sets of requirements for sustainment were identified: 1) characteristics of the provider, including competence in delivering the intervention, need for continued technical support and assistance from outside experts, and satisfaction with addressing the needs of this population; 2) characteristics of the clients (i.e., FSWs), including client need and demand for services and incentives for participation; 3) characteristics of the organization, including its mission, benefits, and operations; 4) characteristics of the outer setting, including financial support and relationship with the community-based organization’s central offices, and transportation and security in areas where FSWs live and work; and 5) outcomes associated with the intervention itself, including a reduction of risk through education and increased outreach through referrals from FSWs who received the intervention.

Conclusions

Although the requirements for successful sustainment of interventions like Mujer Segura are consistent with the factors identified in many models of implementation, the results illustrate the importance of local context in assigning priority to these model elements and suggest that the five categories are not discrete entities but interconnected.     

Prevalence and Correlates of Client-Perpetrated Violence against Female Sex Workers in 13 Mexican Cities

Background

Globally, client-perpetrated violence against female sex workers (FSWs) has been associated with multiple health-related harms, including high-risk sexual behavior and increased exposure to HIV/STIs. This study examined correlates of client-perpetrated sexual, physical, and economic violence (e.g., robbery) against FSWs in 13 cities throughout Mexico.

Methods

FSWs (N = 1,089) who were enrolled in a brief, evidence-based, sexual risk reduction intervention for FSWs (Mujer Segura) were interviewed about their work context, including experiences of violence perpetrated by clients, sexual risk and substance use practices, financial need, and social supports. Three broad categories of factors (sociodemographic, work context, behavioral and social characteristics of FSWs) were examined as correlates of sexual, physical, and economic violence.

Results

The prevalence of different types of client-perpetrated violence against FSWs in the past 6 months was: sexual (11.7%), physical (11.8%), economic (16.9%), and any violence (22.6%). Greater financial need, self-identification as a street worker, and lower perceived emotional support were independently associated with all three types of violence. Alcohol use before or during sex with clients in the past month was associated with physical and sexual violence. Using drugs before or during sex with clients, injection drug use in the past month, and population size of city were associated with sexual violence only, and FSWs’ alcohol use score (AUDIT-C) was associated with economic violence only.

Conclusions

Correlates of client-perpetrated violence encompassed sociodemographic, work context, and behavioral and social factors, suggesting that approaches to violence prevention for FSWs must be multi-dimensional. Prevention could involve teaching FSWs strategies for risk avoidance in the workplace (e.g., avoiding use of alcohol with clients), enhancement of FSWs’ community-based supports, development of interventions that deliver an anti-violence curriculum to clients, and programs to address FSWs’ financial need by increasing their economic opportunities outside of the sex trade.     

Immunoproteomics of the active degradome to identify biomarkers for Trichomonas vaginalis

Trichomonas vaginalis, a sexually transmitted parasite, has many cysteine proteinases (CPs); some are involved in trichomonal pathogenesis, express during infection, and antibodies against CPs have been detected in patient sera. The goal of this study was to identify the antigenic proteinases of T. vaginalis as potential biomarkers for trichomonosis. The proteases detected when T. vaginalis protein extracts are incubated without protease inhibitors, the trichomonad‐active degradome, and the immunoproteome were obtained by using 2‐DE, 2‐D‐zymograms, 2‐D‐Western blot (WB) assays with trichomonosis patient sera, and MS analysis. Forty‐nine silver‐stained spots were detected in the region of 200–21 kDa of parasite protease‐resistant extracts. A similar proteolytic pattern was observed in the 2‐D zymograms. Nine CPs were identified in the 30 kDa region (TvCP1, TvCP2, TvCP3, TvCP4, TvCP4‐like, TvCP12, TvCPT, TvLEGU‐1, and another legumain‐like CP). The major reactive spots to T. vaginalis‐positive patient sera by 2‐D‐WB corresponded to four papain‐like (TvCP2, TvCP4, TvCP4‐like, TvCPT), and one legumain‐like (TvLEGU‐1) CPs. The genes of TvCP4, TvCPT, and TvLEGU‐1 were cloned, sequenced, and expressed in Escherichia coli. Purified recombinant CPs were recognized by culture‐positive patient sera in 1‐D‐WB assays. These data show that some CPs could be potential biomarkers for serodiagnosis of trichomonosis.     

Growth, mortality and reproduction of the blue tilapia Oreochromis aureus (Perciformes: Cichlidae) in the Aguamilpa Reservoir, Mexico

Tilapia production has increased in Aguamilpa Reservoir, in Nayarit, Mexico, in the last few years and represents a good economic activity for rural communities and the country. We determined growth parameters, mortality and reproductive aspects for 2413 specimens of blue tilapia Oreochromis aureus in this reservoir. Samples were taken monthly from July 2000 through June 2001, of which 1 371 were males and 1 042 were females. Standard length (SL) and total weight (TW) were measured in each organism. The SL/TW relationships through power models for sexes were determined. The growth parameters L infinity k, and t0 of the von Bertalanffy equation were estimated using frequency distribution of length through ELEFAN-I computer program. Finally the reproductive cycle and size of first maturity were established using morph chromatic maturity scale. The results suggested that the males and females had negative allometric growth (b < 3). Significant differences were found between SL/TW model for the sexes, suggesting separate models for males and females. Results indicate that there are no differences in growth rates between sexes; the proposed parameters were L infinity = 43.33 cm standard length, k = 0.36/year and t0 = -0.43 years. Natural and fishing mortality coefficients were 0.83/year and 1.10/year, respectively. The estimated exploitation rate (0.57/year) suggested that during the study period the fishery showed signs of overfishing. Blue tilapia reproduces year-round; the highest activity occurs from January through May and size of first maturity was 23 cm SL. We conclude that it is necessary to establish a minimum catch size in this reservoir based on the reproductive behavior of this species.     

Building an understanding of cystic fibrosis on the foundation of ABC transporter structures

Cystic fibrosis (CF) is a fatal disease affecting the lungs and digestive system by impairment of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR). While over 1000 mutations in CFTR have been associated with CF, the majority of cases are linked to the deletion of phenylalanine 508 (ΔF508). F508 is located in the first nucleotide binding domain (NBD1) of CFTR. This mutation is sufficient to impair the trafficking of CFTR to the plasma membrane and, thus, its function. As an ABC transporter, recent structural data from the family provide a framework on which to consider the effect of the ΔF508 mutation on CFTR. There are fifty-seven known structures of ABC transporters and domains thereof. Only six of these structures are of the intact transporters. In addition, modern bioinformatic tools provide a wealth of sequence and structural information on the family. We will review the structural information from the RCSB structure repository and sequence databases of the ABC transporters. The available structural information was used to construct a model for CFTR based on the ABC transporter homologue, Sav1866, and provide a context for understanding the molecular pathology of Cystic Fibrosis.     

Seed-size variation determines interspecific differential predation by mammals in a neotropical rain forest

It has been suggested that the anthropogenically driven loss of herbivorous mammals might lead to changes in the recruitment patterns of tropical rain forest plants, but few studies have examined the explicit mechanisms that might account for this effect. Here we propose a conceptual model linking differential mammalian defaunation and differential plant recruitment. We posit that in the absence of medium/large herbivores but with small rodent granivores still present (i.e. differential defaunation), predation pressure will be greater on small-seeded species than on large-seeded species. We tested such differential predation hypothesis (DPH) by means of a series of experiments directed to assess: 1) seed consumption by small rodents in laboratory cages; 2) seed-removal rates in small rodent enclosures in the field and 3) removal of seeds placed on the forest floor and exposed to either the full complement of mammals or only small rodents. Seeds used in the experiments were arranged in pairs consisting of species from the same taxonomic family but with a contrasting size (large, small). We found: 1) a significantly greater consumption of smaller seeds (a 2.3- to 20.5-fold difference) in cages and 2) a significantly greater removal of small-seeded species (a 3.7- to 65-fold difference) in field enclosures. Results of seed removal experiments in free-access plots and selective exclosures were more complex, with a general absence of significant differences among treatments but we found that predation was in general concentrated on small-seeded species and small rodents were the predominant visitors to the plots. This, together with the overall short distance of dispersal among large seeds suggest that in differentially defaunated forests large-seeded species are more likely to escape predation. We posit that such size related differential predation may lead to the floristic impoverishment observed in some defaunated forests.