Assay strategies for the discovery and validation of therapeutics targeting <i>Brugia pahangi</i> Hsp90

The chemotherapy of lymphatic filariasis relies upon drugs such as diethylcarbamazine and ivermectin that largely target the microfilarial stages of the parasite, necessitating continued treatment over the long reproductive life span of the adult worm. The identification of compounds that target adult worms has been a long-term goal of WHO. Here we describe a fluorescence polarization assay for the identification of compounds that target Hsp90 in adult filarial worms. The assay was originally developed to identify inhibitors of Hsp90 in tumor cells, and relies upon the ability of small molecules to inhibit the binding of fluorescently labelled geldanamycin to Hsp90. We demonstrate that the assay works well with soluble extracts of Brugia, while extracts of the free-living nematode C. elegans fail to bind the probe, in agreement with data from other experiments. The assay was validated using known inhibitors of Hsp90 that compete with geldanamycin for binding to Hsp90, including members of the synthetic purine-scaffold series of compounds. The efficacy of some of these compounds against adult worms was confirmed in vitro. Moreover, the assay is sufficiently sensitive to differentiate between binding of purine-scaffold compounds to human and Brugia Hsp90. The assay is suitable for high-throughput screening and provides the first example of a format with the potential to identify novel inhibitors of Hsp90 in filarial worms and in other parasitic species where Hsp90 may be a target

Hemodialysis in infants and small children

Hemodialysis in infants and small children requires specialized nursing staff, equipment and adequate access. The techniques, requirements and available equipment for this population are discussed.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/47834/1/467_2004_Article_BF00868283.pd

Does Father Know Best? A Formal Model of the Paternal Influence on Childhood Social Anxiety

We explore paternal social anxiety as a specific risk factor for childhood social anxiety in a rational optimization model. In the course of human evolution, fathers specialized in external protection (e.g., confronting the external world) while mothers specialized in internal protection (e.g., providing comfort and food). Thus, children may instinctively be more influenced by the information signaled by paternal versus maternal behavior with respect to potential external threats. As a result, if fathers exhibit social anxiety, children interpret it as a strong negative signal about the external social world and rationally adjust their beliefs, thus becoming stressed. Under the assumption that paternal signals on social threats are more influential, a rational cognitive inference leads children of socially anxious fathers to develop social anxiety, unlike children of socially anxious mothers. We show in the model that mothers cannot easily compensate for anxious paternal behavior, but choose to increase maternal care to maintain the child’s wellbeing. We discuss research directions to test the proposed model as well as implications for the prevention and treatment of child social anxiety

Glutamate-Gated Chloride Channels of Haemonchus contortus Restore Drug Sensitivity to Ivermectin Resistant Caenorhabditis elegans

Anthelmintic resistance is a major problem in livestock farming, especially of small ruminants, but our understanding of it has been limited by the difficulty in carrying out functional genetic studies on parasitic nematodes. An important nematode infecting sheep and goats is Haemonchus contortus; in many parts of the world this species is resistant to almost all the currently available drugs, including ivermectin. It is extremely polymorphic and to date it has proved impossible to relate any sequence polymorphisms to its ivermectin resistance status. Expression of candidate drug-resistance genes in Caenorhabditis elegans could provide a convenient means to study the effects of polymorphisms found in resistant parasites, but may be complicated by differences between the gene families of target and model organisms. We tested this using the glutamate-gated chloride channel (GluCl) gene family, which forms the ivermectin drug target and are candidate resistance genes. We expressed GluCl subunits from C. elegans and H. contortus in a highly resistant triple mutant C. elegans strain (DA1316) under the control of the avr-14 promoter; expression of GFP behind this promoter recapitulated the pattern previously reported for avr-14. Expression of ivermectin-sensitive subunits from both species restored drug sensitivity to transgenic worms, though some quantitative differences were noted between lines. Expression of an ivermectin-insensitive subunit, Hco-GLC-2, had no effect on drug sensitivity. Expression of a previously uncharacterised parasite-specific subunit, Hco-GLC-6, caused the transgenic worms to become ivermectin sensitive, suggesting that this subunit also encodes a GluCl that responds to the drug. These results demonstrate that both orthologous and paralogous subunits from C. elegans and H. contortus are able to rescue the ivermectin sensitivity of mutant C. elegans, though some quantitative differences were observed between transgenic lines in some assays. C. elegans is a suitable system for studying parasitic nematode genes that may be involved in drug resistance

Developmental and evolutionary assumptions in a study about the impact of premature birth and low income on mother–infant interaction

In order to study the impact of premature birth and low income on mother–infant interaction, four Portuguese samples were gathered: full-term, middle-class (n=99); premature, middle-class (n=63); full-term, low income (n=22); and premature, low income (n=21). Infants were filmed in a free play situation with their mothers, and the results were scored using the CARE Index. By means of multinomial regression analysis, social economic status (SES) was found to be the best predictor of maternal sensitivity and infant cooperative behavior within a set of medical and social factors. Contrary to the expectations of the cumulative risk perspective, two factors of risk (premature birth together with low SES) were as negative for mother–infant interaction as low SES solely. In this study, as previous studies have shown, maternal sensitivity and infant cooperative behavior were highly correlated, as was maternal control with infant compliance. Our results further indicate that, when maternal lack of responsiveness is high, the infant displays passive behavior, whereas when the maternal lack of responsiveness is medium, the infant displays difficult behavior. Indeed, our findings suggest that, in these cases, the link between types of maternal and infant interactive behavior is more dependent on the degree of maternal lack of responsiveness than it is on birth status or SES. The results will be discussed under a developmental and evolutionary reasonin

Chronic dialysis in the infant less than 1 year of age

Dialysis in the infant carries a mortality rate of 16%. Institution of dialysis may be the result of adequate nutritional intake, but avoidance of nutritional intake should never be seen as a way to prevent dialysis. Increased caloric intake, usually via enteral feeding tubes, is needed for optimal growth in the infant with end-stage renal disease (ESRD) in order to attain adequate nutrition with resulting good growth. “Renal” formulae may be constituted as dilute (as in the polyuric infant) or concentrated (as in the anuric infant) to fit the infants needs. Peritoneal dialysis (PD) is the usual mode of renal replacement therapy (97%), with access via a surgically placed cuffed catheter with attention to the placement of the exit site in order to avoid fecal or urinary contamination. PD volumes of 30–40 ml/kg per pass or 800–1,200 ml/m 2 per pass usually result in dialysis adequacy. Additional dietary sodium (3–5 mEq/kg per day) and protein (3–4 g/kg per day) are needed, due to sodium and protein losses in the dialysate. Protein losses are associated with significant infectious morbidity and nonresponsiveness to routine immunizations. Hemodialysis (HD) can be performed either as single- or dual-needle access that have minimal dead space (less then 2 ml) and recirculation rate (less then 5%). Attnetion to extracorporeal blood volume (<10% of intravascular volume), blood flow rates (3–5 ml/kg per min), heparinization (activated clotting times), ultrafiltration (ultrafiltration monitor), and temperature control is imperative during each treatment. Because infants' nutrition is mostly fluid, HD may be needed 4–6 days/week (especially in the oligoanuric infant) to avoid excessive volume overload between treatments. At the end of the treatment a slow blood return with minimal saline rinse is needed to avoid hemodynamic compromise. Infant dialysis, although technically challenging with a significant morbidity and mortality rate, can be safely carried out in the infant with ESRD but requires infant-specific equipment and trained personnel.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/47836/1/467_2004_Article_BF00867678.pd

A Program for At-Risk High School Students Informed by Evolutionary Science

Improving the academic performance of at-risk high school students has proven difficult, often calling for an extended day, extended school year, and other expensive measures. Here we report the results of a program for at-risk 9th and 10th graders in Binghamton, New York, called the Regents Academy that takes place during the normal school day and year. The design of the program is informed by the evolutionary dynamics of cooperation and learning, in general and for our species as a unique product of biocultural evolution. Not only did the Regents Academy students outperform their comparison group in a randomized control design, but they performed on a par with the average high school student in Binghamton on state-mandated exams. All students can benefit from the social environment provided for at-risk students at the Regents Academy, which is within the reach of most public school districts