Modeling Hematologic and Biochemical Parameters with Spatiotemporal Analysis for the Free-Ranging Eastern Box Turtle (Terrapene carolina carolina) in Illinois and Tennessee, a Potential Biosentinel.

Box turtles are long-lived, inhabit both aquatic and terrestrial habitats, and have relatively small home ranges making them a suitable candidate as a sentinel. To characterize their changes in response to environmental health, assessment of observed variation of this species is required. Thus, a comparative health assessment was employed in 825 Eastern box turtles in east central Illinois and Oak Ridge, Tennessee, to establish a baseline health assessment with regard to sex, age class, season, and location, identify temporal trends, and map parameters. Hematological and plasma biochemical variables measured included packed cell volume, total solids, white blood cell and differential counts, calcium, phosphorus, aspartate aminotransferase, bile acids, creatine kinase, and uric acid. These variables were reduced to four principle components that explained 68.8 % of the cumulative variance. The top model included the main effects of year, location, and sex, but no interactions. Spatial analysis of turtles in Tennessee demonstrated increased WBC and decreased PCV in 2011 associated with a clear-cut silviculture site. The results of this health assessment can serve as a baseline of population health in future studies and aid in the utility of this species as a sentinel.

Rare mutations in RINT1 predispose carriers to breast and Lynch syndrome-spectrum cancers.

UNLABELLED: Approximately half of the familial aggregation of breast cancer remains unexplained. A multiple-case breast cancer family exome-sequencing study identified three likely pathogenic mutations in RINT1 (NM_021930.4) not present in public sequencing databases: RINT1 c.343C>T (p.Q115X), c.1132_1134del (p.M378del), and c.1207G>T (p.D403Y). On the basis of this finding, a population-based case-control mutation-screening study was conducted that identified 29 carriers of rare (minor allele frequency < 0.5%), likely pathogenic variants: 23 in 1,313 early-onset breast cancer cases and six in 1,123 frequency-matched controls [OR, 3.24; 95% confidence interval (CI), 1.29-8.17; P = 0.013]. RINT1 mutation screening of probands from 798 multiple-case breast cancer families identified four additional carriers of rare genetic variants. Analysis of the incidence of first primary cancers in families of women carrying RINT1 mutations estimated that carriers were at increased risk of Lynch syndrome-spectrum cancers [standardized incidence ratio (SIR), 3.35; 95% CI, 1.7-6.0; P = 0.005], particularly for relatives diagnosed with cancer under the age of 60 years (SIR, 10.9; 95% CI, 4.7-21; P = 0.0003). SIGNIFICANCE: The work described in this study adds RINT1 to the growing list of genes in which rare sequence variants are associated with intermediate levels of breast cancer risk. Given that RINT1 is also associated with a spectrum of cancers with mismatch repair defects, these findings have clinical applications and raise interesting biological questions.

Rare key functional domain missense substitutions in MRE11A, RAD50, and NBN contribute to breast cancer susceptibility: results from a Breast Cancer Family Registry case-control mutation-screening study.

INTRODUCTION: The MRE11A-RAD50-Nibrin (MRN) complex plays several critical roles related to repair of DNA double-strand breaks. Inherited mutations in the three components predispose to genetic instability disorders and the MRN genes have been implicated in breast cancer susceptibility, but the underlying data are not entirely convincing. Here, we address two related questions: (1) are some rare MRN variants intermediate-risk breast cancer susceptibility alleles, and if so (2) do the MRN genes follow a BRCA1/BRCA2 pattern wherein most susceptibility alleles are protein-truncating variants, or do they follow an ATM/CHEK2 pattern wherein half or more of the susceptibility alleles are missense substitutions? METHODS: Using high-resolution melt curve analysis followed by Sanger sequencing, we mutation screened the coding exons and proximal splice junction regions of the MRN genes in 1,313 early-onset breast cancer cases and 1,123 population controls. Rare variants in the three genes were pooled using bioinformatics methods similar to those previously applied to ATM, BRCA1, BRCA2, and CHEK2, and then assessed by logistic regression. RESULTS: Re-analysis of our ATM, BRCA1, and BRCA2 mutation screening data revealed that these genes do not harbor pathogenic alleles (other than modest-risk SNPs) with minor allele frequencies>0.1% in Caucasian Americans, African Americans, or East Asians. Limiting our MRN analyses to variants with allele frequencies of <0.1% and combining protein-truncating variants, likely spliceogenic variants, and key functional domain rare missense substitutions, we found significant evidence that the MRN genes are indeed intermediate-risk breast cancer susceptibility genes (odds ratio (OR)=2.88, P=0.0090). Key domain missense substitutions were more frequent than the truncating variants (24 versus 12 observations) and conferred a slightly higher OR (3.07 versus 2.61) with a lower P value (0.029 versus 0.14). CONCLUSIONS: These data establish that MRE11A, RAD50, and NBN are intermediate-risk breast cancer susceptibility genes. Like ATM and CHEK2, their spectrum of pathogenic variants includes a relatively high proportion of missense substitutions. However, the data neither establish whether variants in each of the three genes are best evaluated under the same analysis model nor achieve clinically actionable classification of individual variants observed in this study.

Association of vaccine-related attitudes and beliefs between parents and health care providers.

OBJECTIVES: Health care providers influence parental vaccination decisions. Over 90% of parents report receiving vaccine information from their child's health care provider. The majority of parents of vaccinated children and children exempt from school immunization requirements report their child's primary provider is a good source for vaccine information. The role of health care providers in influencing parents who refuse vaccines has not been fully explored. The objective of the study was to determine the association between vaccine-related attitudes and beliefs of health care providers and parents. METHODS: We surveyed parents and primary care providers of vaccinated and unvaccinated school age children in four states in 2002-2003 and 2005. We measured key immunization beliefs including perceived risks and benefits of vaccination. Odds ratios for associations between parental and provider responses were calculated using logistic regression. RESULTS: Surveys were completed by 1367 parents (56.1% response rate) and 551 providers (84.3% response rate). Parents with high confidence in vaccine safety were more likely to have providers with similar beliefs, however viewpoints regarding disease susceptibility and severity and vaccine efficacy were not associated. Parents whose providers believed that children get more immunizations than are good for them had 4.6 higher odds of holding that same belief compared to parents whose providers did not have that belief. CONCLUSIONS: The beliefs of children's health care providers and parents, including those regarding vaccine safety, are similar. Provider beliefs may contribute to parental decisions to accept, delay or forgo vaccinations. Parents may selectively choose providers who have similar beliefs to their own.

Are Recent Medical Graduates More Skeptical of Vaccines?

Rates of delay and refusal of recommended childhood vaccines are increasing in many U.S. communities. Children's health care providers have a strong influence on parents' knowledge, attitudes, and beliefs about vaccines. Provider attitudes towards immunizations vary and affect their immunization advocacy. One factor that may contribute to this variability is their familiarity with vaccine-preventable diseases and their sequelae. The purpose of this study was to investigate the association of health care provider year of graduation with vaccines and vaccine-preventable disease beliefs. We conducted a cross sectional survey in 2005 of primary care providers identified by parents of children whose children were fully vaccinated or exempt from one or more school immunization requirements. We examined the association of provider graduation cohort (5 years) with beliefs on immunization, disease susceptibility, disease severity, vaccine safety, and vaccine efficacy. Surveys were completed by 551 providers (84.3% response rate). More recent health care provider graduates had 15% decreased odds of believing vaccines are efficacious compared to graduates from a previous 5 year period; had lower odds of believing that many commonly used childhood vaccines were safe; and 3.7% of recent graduates believed that immunizations do more harm than good. Recent health care provider graduates have a perception of the risk-benefit balance of immunization, which differs from that of their older counterparts. This change has the potential to be reflected in their immunization advocacy and affect parental attitudes.

Immunization safety in US print media, 1995-2005.

OBJECTIVE: To identify and describe vaccine safety in US newspaper articles. METHODS: Articles (1147) from 44 states and Washington, DC, between January 1, 1995, and July 15, 2005, were identified by using the search terms "immunize or vaccine" and "adverse events or safety or exemption or danger or risk or damage or injury or side effect" and were coded by using a standardized data-collection instrument. RESULTS: The mean number of vaccine-safety articles per state was 26. Six (not mutually exclusive) topics were identified: vaccine-safety concerns (46%); vaccine policy (44%); vaccines are safe (20%); immunizations are required (10%); immunizations are not required (8%); and state/school exemption (8%). Three spikes in the number of newspaper articles about vaccine-safety issues were observed: in 1999 regarding rotavirus vaccine and in 2002 and 2003 regarding smallpox vaccine. Excluding articles that referred to rotavirus and smallpox vaccines, 37% of the articles had a negative take-home message. CONCLUSION: Ongoing monitoring of news on vaccine safety may help the content and framing of vaccine-safety messages.

Prenatal immunization education the pediatric prenatal visit and routine obstetric care.

BACKGROUND: Vaccine safety concerns and lack of knowledge regarding vaccines contribute to delays in infant immunization. Prenatal vaccine education could improve risk communication and timely vaccination. This study sought to determine the proportion of obstetric practices and hospital-based prenatal education classes that provide pregnant women with infant immunization information, the willingness of obstetric practices to provide infant immunization information, and the proportion of first-time mothers who receive a pediatric prenatal visit. METHODS: A telephone survey was conducted of 100 pediatric practices and 100 obstetric practices randomly selected from the American Medical Association Physician Masterfile between January and March 2005, with analysis performed April 2005. RESULTS: Seventy-one of 100 (71%) selected obstetric practices and 85 of 100 (85%) selected pediatric practices participated. Sixteen obstetric practices (23%) reported providing pregnant women with information on routine childhood immunizations. Thirty-four of the 52 practices (65%) that did not provide such information reported willingness to do so. Ten of 51 hospitals (20%) did not provide information about routine childhood immunizations to prenatal class participants. Sixty-six of the 85 pediatric practices (78%) provided a pediatric prenatal visit. Among these, the median percentage of first-time mothers who received a visit was 30%. CONCLUSIONS: Prenatal visits are a missed opportunity for providing education about infant immunizations. Incorporating immunization education into routine obstetric prenatal care may increase maternal knowledge of infant vaccines and reduce delayed immunization.

A sex-specific metabolite identified in a marine invertebrate utilizing phosphorus-31 nuclear magnetic resonance.

Hormone level differences are generally accepted as the primary cause for sexual dimorphism in animal and human development. Levels of low molecular weight metabolites also differ between men and women in circulating amino acids, lipids and carbohydrates and within brain tissue. While investigating the metabolism of blue crab tissues using Phosphorus-31 Nuclear Magnetic Resonance, we discovered that only the male blue crab (Callinectes sapidus) contained a phosphorus compound with a chemical shift well separated from the expected phosphate compounds. Spectra obtained from male gills were readily differentiated from female gill spectra. Analysis from six years of data from male and female crabs documented that the sex-specificity of this metabolite was normal for this species. Microscopic analysis of male and female gills found no differences in their gill anatomy or the presence of parasites or bacteria that might produce this phosphorus compound. Analysis of a rare gynandromorph blue crab (laterally, half male and half female) proved that this sex-specificity was an intrinsic biochemical process and was not caused by any variations in the diet or habitat of male versus female crabs. The existence of a sex-specific metabolite is a previously unrecognized, but potentially significant biochemical phenomenon. An entire enzyme system has been synthesized and activated only in one sex. Unless blue crabs are a unique species, sex-specific metabolites are likely to be present in other animals. Would the presence or absence of a sex-specific metabolite affect an animal's development, anatomy and biochemistry?

The cerebroplacental Doppler ratio and neonatal outcome in diamnionic monochorionic and dichorionic twins.

OBJECTIVE: Neonatal outcome in twins was studied in relation to the cerebroplacental ratio (CPR). METHODS: Seventy-five infants from twin pregnancies with fetal Doppler data obtained within 3 weeks of delivery were candidates for study (23 infants from diamnionic monochorionic and 52 infants from diamnionic dichorionic twin pregnancies). Multivariate regression analyses were expanded to include 114 twin infants (34 diamnionic monochorionic and 80 diamnionic dichorionic twins). Patients with twin transfusion syndrome were excluded from analysis in the monochorionic group. Targeted ultrasound examination with biometry was performed, and Doppler resistance index (RI) of the umbilical artery (UA) and the middle cerebral artery (MCA) were obtained, and the CPR, a measure of blood flow redistribution, was calculated. Outcome variables included major complications, growth restriction, days of ventilator and oxygen use, days in the neonatal intensive care unit and length of stay. RESULTS: The CPR was correlated more highly with adverse outcomes such as birth weight, special-care nursery days and length of stay than were the UA RI or the MCA RI. The CPR was significantly lower in monochorionic compared with dichorionic twins (1.12 vs. 1.27, p = 0.01). Multivariate regression analyses conducted separately on each twin group also demonstrated that CPR was superior to UA RI and MCA RI in predicting length of stay and restricted growth. Among the Doppler variables, the CPR showed the highest sensitivity for growth restriction (67%). CONCLUSION: In twins, CPR was superior to UA RI and MCA RI in predicting adverse neonatal events.

Evaluating the value of screening for hypertension: an evidence-based approach.

No recommendations regarding in-school blood pressure (BP) screening currently exist. The purpose of this project was to use an evidence-based approach to determine whether BP screening should be initiated as part of one school district's standard screening protocols. Pediatric BP measurement, risk factors for hypertension, issues for determining youth at risk for hypertension, and eligibility criteria for determining conditions appropriate for screening are discussed. BPs of 1st, 6th, and 11th graders were evaluated according to standardized criteria. The evidence indicated that BP screening in school appears warranted, although a formalized study is needed before a definitive decision can be made regarding the incorporation of BP screening into school health services.

Immunomorphometric studies of proteinuria in individual deep and superficial nephrons of rats.

Heterogeneity of structure and function among nephrons is a well-recognized feature of chronic renal diseases. However, only a small number of superficial nephrons per kidney are accessible for micropuncture analysis and relationships of proteinuria to structural change in individual nephrons of experimental models are not clearly established. To directly evaluate proteinuria in many individual nephrons, we developed an immunomorphometric method of analysis. This method is based on the uniformly abundant renal synthesis of Tamm-Horsfall protein (THP) in the thick ascending limb of Henle's loop (TAL). Luminal rabbit immunoglobulin G (IgG) deposits are formed in TALs of proteinuric nephrons in rats injected with heterologous IgG anti-THP antibodies. This immunomorphometric luminal deposit method of assessing proteinuria was previously validated through analysis of heterologous immune complex nephropathy. Glomerular dysfunction in several models-spontaneously hypertensive rats (SHR), aging Sprague-Dawley (SD) rats, rats with adriamycin nephropathy (ADR), and rats subjected to subtotal nephrectomy (NX)-was characterized by immunomorphometric analysis after injection of anti-THP antibodies. Luminal IgG deposits were used to identify nephrons with increased proteinuria. Nephrons were identified histologically as either long looped (LL) or short looped (SL), and frequency of luminal deposits in these nephrons was determined. Glomerular size and sclerosis in deep and superficial zones of renal cortex were determined. Luminal deposits in LL nephrons were more frequent than luminal deposits in SL nephrons in SHRs (p < .001) and aging rats (p < .001) and SL nephrons in ADR rats (p < .02). Whole kidney levels of albuminuria correlated closely with the frequency of luminal deposits in both LL and SL nephrons of SHRs and ADR rats and in LL nephrons of aging rats (p < .005). In contrast, LL and SL deposits were equal in NX rats and did not correlate with albuminuria. A majority of luminal deposits extended beyond the first medullary TAL zone of NX rats, but was confined to this zone in the other 3 models. Deep cortical glomeruli were larger with more glomerulosclerosis than superficial cortical glomeruli. Albuminuria correlated with sclerosis of both deep (p < .002) and superficial (p < .01) glomeruli in NX rats, but not in the other three models. These studies provide a detailed characterization of a new method that allows comparison of proteinuria derived from deep and superficial nephrons. They also provide evidence that pathogenesis of the glomerulosclerosis in NX rats differs from that of the other three models. Glomerulosclerosis was closely linked to the overall level of albuminuria in NX rats, but not to luminal deposits. In the other three models, albuminuria and luminal deposits were closely linked but did not correlate with glomerulosclerosis. Furthermore, LL and SL nephron proteinuria of NX rats was comparable while LL proteinuria was markedly greater than SL proteinuria in the other three models. The luminal deposit method provides a new way to analyze heterogeneity of proteinuria among nephrons and the mechanisms underlying structural change in experimental glomerular diseases.

My experience with health care reform.

Health care reform efforts have personal and professional implications as well as consequences. This case study describes the author's experience with suddenly losing his chaplaincy position because of budgetary constraints. Methods of coping with the trauma are described.

10-nmol oligonucleotide synthesis for the ABI model 394 DNA synthesizer.

Sequencing and gene contig assembly generally involve primer walking, in which an oligonucleotide primer is used once and then discarded. Because the smallest commonly available scale for commercial oligonucleotide synthesizers is the 40-nmol scale, a large excess of product is produced. This results in reagent waste, excess cost to the researcher, and increased production time. A logical solution would be to develop a smaller scale for the commercial synthesizers. A 10-nmol scale was developed for the Applied Biosystems ABI model 394 DNA synthesizer. To develop the 10-nmol scale, a column support was first developed from commonly available materials. Then a 10-nmol cycle was developed from the systematic reduction of the reagents. The final product of the 10-nmol cycle was tested for quality by monitoring the coupling efficiency, polyacrylamide gel electrophoresis, and automated DNA sequencing.The final cycle reduces the cost of the oligonucleotide and decreases the time required to produce it.

The aminoglycosides.

Despite the introduction of newer, less toxic antimicrobial agents, the aminoglycosides continue to serve a useful role in the treatment of serious enterococcal, mycobacterial, and gram-negative bacillary infections. Gentamicin, because of its low cost, remains the aminoglycoside of choice in hospitals with low levels of resistance among Enterobacteriaceae and Pseudomonas aeruginosa. Typically, it is administered in combination with beta-lactam antibiotics, but it may also be used as monotherapy for urinary tract infections or tularemia. Amikacin is useful against gentamicin-resistant gram-negative bacilli and also in the treatment of infections caused by susceptible Nocardia and nontuberculous mycobacteria. Streptomycin serves an important role in the treatment of multidrug-resistant tuberculosis and may be useful in the treatment of some gentamicin-resistant enterococcal infections. Despite an alarming increase in aminoglycoside-resistant enterococci, most institutions have noted little change in patterns of resistance among gram-negative bacilli. Although the development of newer, less toxic aminoglycosides is unlikely in the near future, single daily dosing regimens have been proposed as a convenient, cost-effective strategy. In selected patients, this novel approach seems to be as safe and effective as traditional, multidose regimens.

Debt management and financial planning support for primary care students and residents at Boston University School of Medicine.

Boston University Medical Center created the Office of Residency Planning and Practice Management as part of The Robert Wood Johnson Foundation's Generalist Physician Initiative. Since 1995, the office has improved the medical center's ability to promote and support the generalist career decisions of its students and residents by removing indebtedness as a disincentive. After a brief review of the relationship between indebtedness and specialty selection, the authors delineate the nature and volume of debt-management assistance provided by the office to students and residents through individual counseling sessions, workshops, and other means between April 1995 and March 1998. A case study shows the progression of these services throughout residency training. The medical center also coordinates its debt-management assistance with counseling from physician-oriented financial planning groups. In conclusion, the authors discuss several characteristics of a successful debt-management program for residents.

Antifungal agents. Part II. The azoles.

Before 1978, amphotericin B and flucytosine were the only drugs available for the treatment of systemic fungal infections. The imidazoles, miconazole and ketoconazole, were introduced during the next 3 years. Intravenously administered miconazole served a limited therapeutic role and is no longer available. Orally administered ketoconazole, an inexpensive, effective, and convenient option for treating mucosal candidiasis, was widely used for a decade because it was the only available oral therapy for systemic fungal infections. During the 1990s, use of ketoconazole diminished because of the release of the triazoles--fluconazole and itraconazole. Fluconazole is less toxic and has several pharmacologic advantages over ketoconazole, including penetration into the cerebrospinal fluid. In addition, it has superior efficacy against systemic candidiasis, cryptococcosis, and coccidioidomycosis. Despite a myriad of drug interactions and less favorable pharmacologic and toxicity profiles in comparison with fluconazole, itraconazole has become a valuable addition to the antifungal armamentarium. It has excellent activity against sporotrichosis and seems promising in the treatment of aspergillosis. Itraconazole has replaced ketoconazole as the therapy of choice for nonmeningeal, non-life-threatening cases of histoplasmosis, blastomycosis, and paracoccidioidomycosis and is effective in patients with cryptococcosis and coccidioidomycosis, including those with meningitis. Further investigation into the development of new antifungal agents is ongoing.

Using narratives to evaluate psychiatry residents' competence.

The authors propose a new way to assess the clinical competence of psychiatric residents. Faculty eualuators would be trained by using narrative descriptions of appropriate and inappropriate practice in various clinical settings. The training has the potential to provide more realistic data about the clinical abilities of residents than current methods.

The purification, characterization, and primary structure of a small redox protein from Methanobacterium thermoautotrophicum, an archaebacterium.

A small redox-active protein has been purified to homogeneity from cell-free extracts of the strictly anaerobic thermophilic methanogen, Methanobacterium thermoautotrophicum (strain Marburg). The purification consisted of streptomycin sulfate and acid treatments and three chromatographic steps using Sephadex G-75, Mono Q HR 10/10, and Superose 12 HR 10/30 columns. When these procedures were carried out under strictly anaerobic conditions, approximately 3 mg of this protein could be isolated from 45 g of wet cell paste. Like the thioredoxins and glutaredoxins, it is a small acidic protein (pI = 4.2) consisting of 83 amino acids (M(r) = 9136). In the presence of dithiothreitol or dihydrolipoate, the protein serves as a hydrogen donor for the ribonucleotide reductase from Escherichia coli, and it catalyzes the reduction of insulin. However, it does not interact with the thioredoxin reductases from E. coli or Corynebacterium nephridii and does not function as a hydrogen donor for the ribonucleotide reductase of C. nephridii. The amino acid sequences determined by automated Edman degradation of the 14C-carboxymethylated protein and of peptides derived from trypsin and chymotrypsin digestions show a redox-active site -Cys-Pro-Tyr-Cys-, typical of the glutaredoxins. Its amino acid sequence shows moderate identity with the known glutaredoxins (E. coli, yeast, rabbit bone marrow, calf thymus, and pig liver) when the proteins are aligned at the active site. The secondary structure of the glutaredoxin-like protein predicted by the Chou-Fasman procedure shows that it is similar to the known glutaredoxins. However, surprisingly, the protein does not function as a glutathione-disulfide oxidoreductase in the presence of glutathione and glutathione reductase. This glutaredoxin-like protein may be a component of a ribonucleotide-reducing system distinct from the previously described systems utilizing thioredoxin or glutaredoxin.