NPHS2 variation in focal and segmental glomerulosclerosis

Background: Focal and segmental glomerulosclerosis (FSGS) is the most common histologic pattern of renal injury seen in adults with idiopathic proteinuria. Homozygous or compound heterozygous mutations in the podocin gene NPHS2 are found in 10–30% of pediatric cases of steroid resistant nephrosis and/or FSGS. Methods: We studied the spectrum of genetic variation in 371 individuals with predominantly late onset FSGS (mean age of onset 25 years) by analysis of DNA samples. Results: We identified 15 non-synonymous alleles that changed the amino acid sequence in 63 of the subjects screened (17%). Eight of these (p.R138Q, p.V180M, p.R229Q, p.E237Q, p.A242V, p.A284V, p.L327F and the frameshift 855–856 delAA) are alleles previously reported to cause FSGS in either the homozygous or compound heterozygous states, while the remaining 7 (p.R10T, p.V127W, p.Q215X, p.T232I, p.L270F, p.L312V and the frameshift 397delA) are novel alleles that have not been demonstrated previously. Twelve individuals of the 371 (3.2%) screened had two likely disease-causing NPHS2 alleles, present in either a homozygous or compound heterozygous state. We genotyped the two most common of the non-synonymous NPHS2 alleles (p.A242V and p.R229Q) identified by resequencing in participants from the Nurses' Health Study and also genotyped p.R229Q in 3 diabetic cohorts. We found that the presence of either of these variants does not significantly alter the risk of albuminuria in the Nurses' Health participants, nor does p.R229Q associate with "diabetic nephropathy". Conclusion: NPHS2 mutations are a rare cause of FSGS in adults. The most common non-synonymous NPHS2 variants, p.R229Q and p.A242V, do not appear to alter the risk of proteinuria in the general population nor does p.R229Q associate with measures of kidney dysfunction in diabetic individuals. Our results help clarify the frequency of FSGS-causing NPHS2 mutations in adults and broaden our understanding of the spectrum of NPHS2 mutations that lead to human disease

Comparing United States and Canadian population exposures from National Biomonitoring Surveys: Bisphenol A intake as a case study

The Centers for Disease Control and Prevention provides biomonitoring data in the United States as part of the National Health and Nutrition Examination Survey (NHANES). Recently, Statistics Canada initiated a similar survey — the Canadian Health Measures Survey (CHMS). Comparison of US and Canadian biomonitoring data can generate hypotheses regarding human exposures from environmental media and consumer products. To ensure that such comparisons are scientifically meaningful, it is essential to first evaluate aspects of the surveys' methods that can impact comparability of data. We examined CHMS and NHANES methodologies, using bisphenol A (BPA) as a case study, to evaluate whether survey differences exist that would hinder our ability to compare chemical concentrations between countries. We explored methods associated with participant selection, urine sampling, and analytical methods. BPA intakes were also estimated to address body weight differences between countries. Differences in survey methods were identified but are unlikely to have substantial impacts on inter-survey comparisons of BPA intakes. BPA intakes for both countries are below health-based guidance values set by the US, Canada and the European Food Safety Authority. We recommend that before comparing biomonitoring data between surveys, a thorough review of methodologic aspects that might impact biomonitoring results be conducted

Hershey Medical Center Technical Workshop Report: Optimizing the design and interpretation of epidemiologic studies for assessing neurodevelopmental effects from in utero chemical exposure

Neurodevelopmental disabilities affect 3-8% of the 4 million babies born each year in the U.S. alone, with known etiology for less than 25% of those disabilities. Numerous investigations have sought to determine the role of environmental exposures in the etiology of a variety of human neurodevelopmental disorders (e.g., learning disabilities, attention deficit-hyperactivity disorder, intellectual disabilities) that are manifested in childhood, adolescence, and young adulthood. A comprehensive critical examination and discussion of the various methodologies commonly used in investigations is needed. The Hershey Medical Center Technical Workshop: Optimizing the design and interpretation of epidemiologic studies for assessing neurodevelopmental effects from in utero chemical exposure provided such a forum for examining these methodologies. The objective of the Workshop was to develop scientific consensus on the key principles and considerations for optimizing the design and interpretation of epidemiologic studies of in utero exposure to environmental chemicals and subsequent neurodevelopmental effects. (The Panel recognized that the nervous system develops post-natally and that critical periods of exposure can span several developmental life stages.) Discussions from the Workshop Panel generated 17 summary points representing key tenets of work in this field. These points stressed the importance of: a well-defined, biologically plausible hypothesis as the foundation of in utero studies for assessing neurodevelopmental outcomes; understanding of the exposure to the environmental chemical(s) of interest, underlying mechanisms of toxicity, and anticipated outcomes; the use of a prospective, longitudinal cohort design that, when possible, runs for periods of 2-5 years, and possibly even longer, in an effort to assess functions at key developmental epochs; measuring potentially confounding variables at regular, fixed time intervals; including measures of specific cognitive and social-emotional domains along with non-cognitive competence in young children, as well as comprehensive measures of health; consistency of research design protocols across studies (i.e., tests, covariates, and analysis styles) in an effort to improve interstudy comparisons; emphasis on design features that minimize introduction of systematic error at all stages of investigation: participant selection, data collection and analysis, and interpretation of results; these would include (but not be limited to) reducing selection bias, using double-blind designs, and avoiding post hoc formulation of hypotheses; a priori data analysis strategies tied to hypotheses and the overall research design, particularly for methods used to characterize and address confounders in any neurodevelopmental study; actual quantitative measurements of exposure, even if indirect, rather than methods based on subject recall; careful examination of standard test batteries to ensure that the battery is tailored to the age group as well as what is known about the specific neurotoxic effects on the developing nervous system; establishment of a system for neurodevelopmental surveillance for tracking the outcomes from in utero exposure across early developmental time periods to determine whether central nervous system injuries may be lying silent until developmentally challenged; ongoing exploration of computerized measures that are culturally and linguistically sensitive, and span the age range from birth into the adolescent years; routine incorporation of narrative in manuscripts concerning the possibility of spurious (i.e., false positive and false negative) test results in all research reportage (this can be facilitated by detailed, transparent reporting of design, covariates, and analyses so that others can attempt to replicate the study); forthright, disciplined, and intellectually honest treatment of the extent to which results of any study are conclusive--that is, how generalizable the results of the study are in terms of the implications for the individual study participants, the community studied, and human health overall; confinement of reporting to the actual research questions, how they were tested, and what the study found, and avoiding, or at least keeping to a minimum, any opinions or speculation concerning public health implications; education of clinicians and policymakers to critically read scientific reports, and to interpret study findings and conclusions appropriately; and recognition by investigators of their ethical duty to report negative as well as positive findings, and the importance of neither minimizing nor exaggerating these findings

Secondary metabolites of bacteria obtained from the northeastern Pacific ocean : structure elucidation and biosynthetic studies

Investigation of the organic extracts obtained from cultures of four species of bacteria isolated from the northeastern Pacific ocean led to the isolation of eight new and six previously known secondary metabolites. The structures of the new compounds were elucidated by extensive spectroscopic analysis. In addition, the biogenetic origins of the atoms in two of the bacterial metabolites were probed using stable isotope incorporation experiments. A culture of the bacterium Serratia odorifera, isolated from a surface water sample taken near a Chinook salmon (Oncorhyncus tshawytscha) farm in Georgia Strait, British Columbia, produced the novel compound oncorhyncolide (34). Oncorhyncolide has a unique structure that is apparently not related to other known microbial metabolites isolated from terrestrial sources. Biosynthetic studies using stable isotopes have shown that all the carbons in oncorhyncolide are derived from acetate. The methyl branches in oncorhyncolide are derived from the C2 of acetate and are attached to carbon atoms derived from the carbonyl carbon, C1, of an acetate unit. This type of methyl branching is very rare in polyketide biosynthesis. Examination of the ethyl acetate extracts of a solid agar culture of Pseudomonas fluorescens obtained from an unidentified tunicate in Moira Sound, Alaska, led to the isolation of moiramides A (38), B (39) and C (40) as well as the known compound andrimid (41). The crude extract had shown antibacterial activity against Staphylococcus aureus and methicillinresistant S. aureus. Moiramide B and andrimid proved to be the compounds responsible for this activity. Biosynthetic studies on andrimid have shown that the acylsuccinimide ring is derived from valine, glycine and acetate. It has been proposed that the biosynthesis proceeds through a dipeptide-like intermediate formed from γ-amino-β-keto acids that are in turn formed from valine and glycine homologated with acetate, presumably via malonyl-CoA. Liquid cultures of Pseudomonas sp. 91V47 obtained from an abalone collected off Cortez Island in Georgia Strait, British Columbia, gave an extract that exhibited potent in vitro cytotoxicity. Bioassay-guided fractionation of the crude extract led to the isolation of three new δ-hydroxy acid rhizoxin analogs (44 to 46). The three new compounds showed significant in vitro activity against P388 murine leukemia. A marine isolate of the bacterium Bacillus pumilus, obtained from a sediment sample collected in Georgia Strait, British Columbia, produced an extract that exhibited antimicrobial activity against Staphylococcus aureus, methicillin-resistant S. aureus and S. saprophytics. The known compound AI-77-B (54), previously isolated from terrestrial and marine sources of B. pumilus, was found to be responsible for the antibacterial activity in the crude extract. A new AI-77-B analog, compound 57, was also isolated. Compound 57 showed antibacterial activity only at high concentrations. [chemical compound diagrams]Science, Faculty ofChemistry, Department ofGraduat

Adsorption of bovine serum albumin to polyethylene tubing reversibility and pH-dependence

This thesis is concerned with the adsorption of bovine serum albumin to polyethylene tubing. A method using radioiodinated protein was developed to measure the surface concentration taking into account the dilution effect for miscible displacement in a capillary. A steady-state surface concentration was established within 2 hours. Adsorption did not depend on the ratio of radiolabelled to unlabel led protein. The adsorption isotherm was Langmuir-like with a plateau concentration of approximately 0.2 μg/cm². Two methods were used to calculate the surface concentration in the desorption study. The surface concentration calculated by depletion of the total radioactivity was always higher than that calculated from assaying the radioactivity associated with the tubing. Desorption of at least 5% of the loosely bound protein occurs. The surface concentration-pH data show two maxima. The first is at the isoelectric point of the albumin while the second is at pH 9.5-10. The second maximum seems to be due to preferential adsorption of the higher molecular weight oligomers in the protein sample.Science, Faculty ofChemistry, Department ofGraduat

Clinical leadership : the challenge for senior leaders

Doctors in senior positions in health care and health-care education organizations are increasingly required to take wider management and leadership responsibilities. A range of development programmes are available to support these leaders at local, regional, national and UK levels

Patient and Patient Group Engagement in Cancer Clinical Trials: A Stakeholder Charter

Background—to guide the implementation of patient centricity and engagement in cancer clinical trials (CTs) and to operationalize the Canadianized version of the Clinical Trials Transformation Initiative (C-CTTI) model, the development of a charter was identified by cancer CT stakeholders. Methods—the Canadian Cancer Trial Stakeholder Charter (the Charter) was initiated by Colorectal Cancer Canada (CCC) and developed via the—1—formation of an inclusive working group (WG) that drafted the document using recommendations collected during the development of the C-CTTI model; 2—socialization of the draft Charter to solicit feedback from cancer CT stakeholders, including those who attended the 2019 CCC Conference; and 3—incorporation of stakeholders’ feedback and finalization of the Charter by the WG. Results—the Charter was built around five guiding principles—1—patient centricity; 2—commitment to education and training; 3—collaboration as equal and independent partners in research; 4—transparency and accountability; and 5—high standards in data collection integrity and honesty. These principles led to the Charter’s five tenets, which stipulate stakeholder commitments, aiming to make CTs accessible to all patients, improve the design and implementation of CTs to benefit patients, expand recruitment and retention of patients in CTs, and further advance cancer research and treatment. Conclusions—the Charter is intended to integrate the patient voice into the Canadian cancer CT continuum. The next phases of the C-CTTI model include the adoption and implementation of the Charter, the establishment of a patient group training program, and the development of real-world evidence/real-world data methodologies