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Determination of the aquatic PNEC0.05 for boron.

Several weight of the evidence approaches for the effects assessment of boron to aquatic organisms have been conducted. The focal point for all of these assessments is the appropriate interpretation of the LC1 for rainbow trout (Oncorhynchus mykiss) from W.J. Birge, J.A. Black, Report No. EPA5601176408. Environmental Protection Agency, Office of Toxic Substances, Washington, DC, 1977, as the LC1 from their studies provided the lowest chronic toxicity values for boron. Other studies by these investigators (W.J. Birge, J.A. Black, Completion Report prepared for the Procter & Gamble Company, Cincinnati, USA, 1981; W.J. Birge, J.A. Black, A.G. Westermann, T.M. Short, S.B. Taylor, M.C. Parekh. Completion Report prepared for the Procter & Gamble Company, Cincinnati, USA, 1984; J.A. Black, J.B. Barnum, W.J. Birge, Chemosphere 26 (1993) 1383-1413) further indicate this species to be particularly sensitive to boron. While weight of the evidence approaches have been necessary for diverse risk assessment needs, they each suffer from potential bias via professional judgment. In order to allay these issues and to update the effects assessment for boron, a probabilistic approach was used. Raw data from the Birge and Black studies were reanalyzed to generate chronic values representative of the adverse effects threshold (e.g., LC10) for several species. A comprehensive list of chronic data from these studies and other studies were combined into a cumulative probability term, allowing for the determination of the fifth percentile protection value, or PNEC0.05. The PNEC0.05 for boron was determined to be at least 1.3 mg B/l.

Animals↗

Glycemic control predicts diabetic extrarenal microvascular complications but not renal survival in patients with moderate to severe chronic kidney disease.

BACKGROUND: Control of blood pressure (BP) and blood glucose can slow the development of diabetic nephropathy (DN). However, BP control may be of relatively more importance than glycemic control on the progression of DN. OBJECTIVE: To determine the effects of glycemic control on renal survival in a predominantly African American diabetic population with moderate-to-severe renal disease. DESIGN: This was a retrospective chart review of all diabetic patients seen in an academic nephrology clinic in 2001 and 2002 for renal survival and its predictors and micro/macrovascular disease. The weighted mean glycosylated hemoglobin (GHb) over followup was determined. Mean GHb < or = 9 was defined as low, and GHb >9 was high. The effect of glycemic control on endpoints was determined by Cox proportional hazards and logistic regression. RESULTS: One hundred fifty-five diabetic patients (87.7% African American, mean creatinine =2.2 mg/dL) had sufficient GHb measurements. Compared to the high group (n=81), the low group (n=74) was significantly younger, had a shorter duration of diabetes, and worse renal function. No significant association of glycemic control with renal survival (ESRD) was seen. Glycemic control and the presence of DN were significantly related to extrarenal microvascular complications, independent of other factors. CONCLUSION: Glycosylated hemoglobin (GHb) is not a significant predictor of renal survival in patients with diabetes and moderate renal disease. However, glycemic control does predict extrarenal microvascular complications in this population. Therefore, good metabolic control remains important in patients with diabetes and renal disease.

Black or African American↗

Improved colorimetric assay for detecting influenza B virus neutralizing antibody responses to vaccination and infection.

An automated neutralization test for influenza B virus is described in which antibody titers are determined according to the release of neutral red from infected or uninfected cells of the Madin-Darby canine kidney line. Endpoints are determined in a standard enzyme-linked immunosorbent assay reader. The test requires no expensive immunologic reagents and was used to evaluate responses to both vaccination and natural infection against influenza B virus. Overall responses to vaccination were comparable with those obtained by hemagglutination inhibition, using Tween-ether-split influenza B/Ann Arbor/1/86 virus as the antigen (the HI-TE test). The sensitivities of neutralization responses compared with those obtained by the HI-TE test for two vaccines were 88 and 89%; the specificities were lower at 61 and 60%, respectively. Responses to vaccination, measured by hemagglutination inhibition, were significantly higher with split virus compared with whole virus. However, seroconversion by both the HI-TE and neutralization tests was observed in 5 of 10 individuals from whom virus was detected by either culture of nasal or throat washings or the presence of antigen from immunofluorescence in cells from nasal washings.

Animals↗

Comparative evaluation of alternative methods for broth dilution susceptibility testing of fluconazole against Candida albicans.

A comparative evaluation of methods for broth macro- and microdilution susceptibility testing of fluconazole was conducted with 119 clinical isolates of Candida albicans. Macro- and microdilution testing were performed according to National Committee for Clinical Laboratory Standards recommendations. For reference macrodilution testing, an 80% inhibition endpoint (MIC 80%) was determined after 48 h of incubation in accordance with National Committee for Clinical Laboratory Standards proposed standard M27-P. Microdilution endpoints were scored as the first tube or well in which a prominent reduction in turbidity (score 2 out of a possible 4) was observed compared with the growth control (Micro MIC-2). Alternative endpoint criteria were assessed independently of the reference MIC 80% and Micro MIC-2 values and included a colorimetric microdilution endpoint determined by using an oxidation-reduction indicator (Alamar Blue; Alamar Bio-sciences Inc., Sacramento, Calif.). The MICs for the two microdilution test systems were read after 24 and 48 h of incubation. The percentage of fluconazole MICs within 2 doubling dilutions of the macrodilution reference values was 94% for both microdilution tests read at 24 h. Agreement was slightly lower at 48 h and ranged from 91 to 93%. Comparison of Micro MIC-2 and colorimetric microdilution MICs resulted in agreements of 97 and 93% at 24 and 48 h, respectively. These results show excellent agreement among alternative methods for fluconazole susceptibility testing.

Candida albicans↗

Phenytoin teratogenicity and midgestational pharmacokinetics in mice.

Mice of the A/J and C57BL/6J (C57) strains were dosed with phenytoin (PHT) every 48 hr throughout pregnancy by gastric intubation to test the hypothesis that maternal plasma PHT concentration may be the significant factor in determining PHT reproductive and developmental toxicity. Serial serum samples were obtained from each mouse from gestation day (GD) 10-GD 12 for determination of individual dam PHT pharmacokinetics. Maximum PHT concentration and PHT AUC (area under-the-time-concentration curve) were regressed to laparotomy and fetal evaluation endpoints to determine whether significant association existed. Although serum PHT concentrations exceeded levels associated with teratogenicity (greater than 10 micrograms/ml), few major malformations were induced in either strain. However, in the A/J strain, there was a significant increased incidence of hydrocephaly and open eyelid. Regression of pharmacokinetic parameters with embryo and maternal endpoints indicated significant associations between gestational weight gain and maximum concentration measured (Cmax) or AUC in both strains. This association was also found for fetal weight in the C57 strain. In the A/J strain, the induction of decreased ossification of the sternebrae was also associated with maternal PHT concentration; however, linear regression of hydrocephaly and open eyelid to PHT concentration was not statistically significant. These results suggest that maternal plasma PHT concentration may be a quantifiable determinant of certain aspects of PHT developmental toxicity in the mouse.

Abnormalities, Drug-Induced↗

Clinical significance of immunohistochemical expression of hypoxia-inducible factor-1alpha as a prognostic marker in rectal adenocarcinoma.

BACKGROUND: Hypoxia-inducible factor-1alpha (HIF-1alpha), a subunit of hypoxia-inducible factor-1 (HIF-1), furnishes tumor cells with the means of adapting to stress parameters, such as tumor hypoxia, and promotes critical steps in tumor progression and aggressiveness by inducing angiogenesis and regulating energy metabolism. In this study, we investigated the relationship between HIF-1alpha and vascular endothelial growth factor (VEGF) and clinicopathologic characteristics, and evaluated the role of HIF-1alpha expression in patients with rectal adenocarcinoma. PATIENTS AND METHODS: The immunohistochemical expression of HIF-1alpha and VEGF was evaluated in 30 formalin-fixed, paraffin-embedded postoperative rectal adenocarcinoma tissue samples. Correlations with clinicopathologic characteristics were determined by cross-tabulations. The impact of the immunoreactivity of HIF-1alpha with regard to the overall survival and local control endpoints was determined by univariate analyses. RESULTS: Increased HIF-1alpha expression was strongly associated with VEGF positivity (P = 0.002), Dukes stage (P = 0.017), and lymph node metastasis (P = 0.001). No correlation was found between the level of HIF-1alpha expression and histologic grade (P = 0.63). The Kaplan-Meier curves showed a significantly shorter overall survival (P = 0.0087) and local control (P = 0.0438) for patients with high HIF-1alpha expression. CONCLUSION: These results show that HIF-1alpha might represent an important biologic marker evaluating the prognosis of patients with rectal adenocarcinoma.

Adenocarcinoma↗

A randomized prospective trial of ioxaglate 320 (Hexabrix) vs. iodixanol 320 (Visipaque) in patients undergoing percutaneous coronary intervention.

We performed a randomized, prospective, double blind trial comparing the use of the ionic dimer contrast agent ioxaglate 320 (Hexabrix) with the nonionic dimer contrast agent iodixanol 320 (Visipaque) in 618 patients undergoing percutaneous coronary intervention (PCI) for stable or unstable coronary artery syndromes. The aim was to determine whether the different anticoagulant and antiplatelet properties of these two contrast agents resulted in a significant difference in the incidence of a combined endpoint comprising the major complications of PCI. Procedural success rates were marginally higher in the Visipaque group compared to the Hexabrix group, although this did not reach statistical significance (96.7% vs. 93.9%; P = 0.09). There was a borderline statistically significant higher requirement for bailout stenting in the Visipaque group compared to the Hexabrix group (6.8% vs. 3.2%; P = 0.05), although this was not a predefined endpoint. The incidence of the combined primary endpoint of failed catheter laboratory outcome/requirement for bailout stenting/requirement for abciximab/myocardial infarction/death before hospital discharge was higher in the Visipaque group compared to the Hexabrix group (17.9% vs. 14.8%), although this did not reach statistical significance (P = 0.29). When subgroup analysis was performed, the incidence of the combined endpoint in patients with stable coronary artery disease randomized to receive either Visipaque or Hexabrix was identical (13.7%). In patients with an acute coronary syndrome, there was a trend toward a reduced incidence of the combined endpoint in the Hexabrix compared to the Visipaque group, although this did not reach statistical significance (17.2% vs. 24.8%; P = 0.17). More adverse reactions occurred in the Hexabrix group compared to the Visipaque group (8.7% vs. 4.9%; P = 0.06). We conclude that there is no clear advantage with the use of an ionic contrast agent in a large population of patients undergoing PCI for both stable and unstable coronary artery disease. Although the study was underpowered to detect significant differences with the use of either agent when patients with either stable or unstable coronary disease were studied, it is highly unlikely that the ionicity of the contrast agent confers any advantage for patients with stable coronary disease. There remains a possibility that ionic agents do have advantages for patients with unstable coronary artery disease undergoing PCI, although a larger study than ours would be required to confirm or refute this.

Adult↗

Circadian rhythm of the PRL response to TRH in healthy subjects.

The PRL response to TRH constitutes an important clinical tool for diagnosing forms of hyperprolactinemic syndrome. Hence it is important to establish the characteristics of the circadian variation in the response of PRL to TRH to improve the diagnostic value of the test. Six male subjects, ranging in age from 23 to 24 years, participated in this study. All were considered healthy on the basis of clinical examination, biochemical and hormonal tests. Six TRH tests were performed on each subject, one test every other day during a total span of 12 days. Each test was performed at a different clock hour: 0000, 0400, 0800, 1200, 1600, 2000. For the test, subjects received 200 microgram TRH intravenously. Blood samples were drawn from a catheterized arm vein before the TRH injection (basal value) and 20, 30, 60 and 120 min after injection. At each timepoint 5 endpoints were determined for PRL on each subject. The population mean cosinor, according to Halberg, was used to investigate the circadian rhythm in each of the endpoints. All the 5 endpoints for PRL are consistent on showing p values near 0.5 and acrophase estimates before midnight (while basal value displays acrophase at 0400). Further investigations are necessary to clarify these circadian rhythms and the shift of the acrophases.

Adult↗

Reproduction disturbances of Brachionus calyciflorus (rotifer) for the screening of environmental endocrine disrupters.

Rotifers and especially the Brachionus calyciflorus species have already shown advantages (e.g. good sensitivity, simplicity, ecological relevance, short generation time) for the chronic toxicity assessment of water pollutants. In this study, the B. calyciflorus three-day test was evaluated on ethinylestradiol, nonylphenol and testosterone for the determination of endocrine disrupting (reproductive) effects. Reproduction (intrinsic rate of population increase) and sex ratio (males/females and mictic females/amictic females) were explored and discussed as suitable endpoints. B. calyciflorus has shown a fair sensitivity to detect reproductive disrupting effects after exposure to ethinylestradiol, nonylphenol and testosterone. Population growth (intrinsic rate of population increase, r) and the ratio ovigerous females/ non-ovigerous females are both suitable endpoints, whereas the occurrence of males was not frequent enough to be used as endpoint.

Animals↗

Microarray analysis of mouse ear tissue exposed to bis-(2-chloroethyl) sulfide: gene expression profiles correlate with treatment efficacy and an established clinical endpoint.

Bis-(2-chloroethyl) sulfide (sulfur mustard; SM) is a potent alkylating agent. Three treatment compounds have been shown to limit SM damage in the mouse ear vesicant model: dimercaprol, octyl homovanillamide, and indomethacin. Microarrays were used to determine gene expression profiles of biopsies taken from mouse ears after exposure to SM in the presence or absence of treatment compounds. Mouse ears were topically exposed to SM alone or were pretreated for 15 min with a treatment compound and then exposed to SM. Ear tissue was harvested 24 h after exposure for ear weight determination, the endpoint used to evaluate treatment compound efficacy. RNA extracted from the tissues was used to generate microarray probes for gene expression profiling of therapeutic responses. Principal component analysis of the gene expression data revealed partitioning of the samples based on treatment compound and SM exposure. Patterns of gene responses to the treatment compounds were indicative of exposure condition and were phenotypically anchored to ear weight. Pretreatment with indomethacin, the least effective treatment compound, produced ear weights close to those treated with SM alone. Ear weights from animals pretreated with dimercaprol or octyl homovanillamide were more closely associated with exposure to vehicle alone. Correlation coefficients between gene expression level and ear weight revealed genes involved in mediating responses to both SM exposure and treatment compounds. These data provide a basis for elucidating the mechanisms of response to SM and drug treatment and also provide a basis for developing strategies to accelerate development of effective SM medical countermeasures.

Administration, Topical↗

Cell-wall-defective variants of Fusobacterium.

The activity of antimicrobial agents against Fusobacterium species has been reported as variable in the literature. For some strains, the inconsistency arises from difficulty in determining the endpoint of growth in agar dilution susceptibility tests. Certain strains persist as a subtle haze beyond the levels of antibiotic that permit conventional colonial growth. We have determined by light and electron microscopy that this haze represents the colonial growth of cell-wall-defective (CWD) variants of the parent Fusobacterium. The CWD forms could be propagated indefinitely in hypertonic medium containing the antibiotic inducing agent. However, when the antibiotic was eliminated, the organisms would revert to their native morphology. Formation of CWD variants was observed in the presence of cell-wall-active drugs (e.g., beta-lactam agents) but not with drugs that work by a different mechanism (e.g., clindamycin or chloramphenicol). Fourteen of 22 F. varium strains, 8 of 11 F. mortiferum strains, 2 of 10 F. gonidiaformans strains, and 1 of 4 of F. necrophorum strains could be induced to a CWD form in vitro in the usual agar dilution susceptibility test. Although the clinical significance of CWD variants of Fusobacterium is unknown, they may be a source of confusion in interpreting agar dilution susceptibility tests.

Anti-Bacterial Agents↗

Interlaboratory evaluation of Etest method for testing antifungal susceptibilities of pathogenic yeasts to five antifungal agents by using Casitone agar and solidified RPMI 1640 medium with 2% glucose.

An interlaboratory evaluation (two centers) of the Etest method was conducted for testing the antifungal susceptibilities of yeasts. The MICs of amphotericin B, fluconazole, flucytosine, itraconazole, and ketoconazole were determined for 83 isolates of Candida spp., Cryptococcus neoformans, and Torulopsis glabrata. Two buffered (phosphate buffer) culture media were evaluated: solidified RPMI 1640 medium with 2% glucose and Casitone agar. MIC endpoints were determined after both 24 and 48 h of incubation at 35 degrees C. Analysis of 3,420 MICs demonstrated higher interlaboratory agreement (percentage of MIC pairs within a 2-dilution range) with Casitone medium than with RPMI 1640 medium when testing amphotericin B (84 to 90% versus 1 to 4%), itraconazole (87% versus 63 to 74%), and ketoconazole (94 to 96% versus 88 to 90%). In contrast, better interlaboratory reproducibility was determined between fluconazole MIC pairs when RPMI 1640 medium rather than Casitone medium was used (96 to 98% versus 77 to 90%). Comparison of the flucytosine MICs obtained with RPMI 1640 medium revealed greater than 80% reproducibility. The study suggests the potential value of the Etest as a convenient alternative method for testing the susceptibilities of yeasts. It also indicates the need for further optimization of medium formulations and MIC endpoint criteria to improve interlaboratory agreement.

Agar↗

A combination of distribution- and anchor-based approaches determined minimally important differences (MIDs) for four endpoints in a breast cancer scale.

OBJECTIVE: To determine distribution- and anchor-based minimal important difference (MID) estimates for four scores from the Functional Assessment of Cancer Therapy-Breast (FACT-B): the breast cancer subscale (BCS), Trial Outcome Index (TOI), FACT-G (the general version), and FACT-B. STUDY DESIGN AND SETTING: We used data from a Phase III clinical trial in metastatic breast cancer (ECOG study 1193; n=739) and a prospective observational study of pain in metastatic breast cancer (n=129). One third and one half of the standard deviation and 1 standard error of measurement were used as distribution-based criteria. Clinical indicators used to determine anchor-based differences included ECOG performance status, current pain, and response to treatment. RESULTS: FACT-B scores were responsive to performance status and pain anchors, but not to treatment response. By combining the results of distribution- and anchor-based methods, MID estimates were obtained: BCS=2-3 points, TOI=5-6 points, FACT-G=5-6 points, and FACT-B=7-8 points. CONCLUSION: Distribution- and anchor-based estimates of the MID do show convergence. These estimates can be used in combination with other measures of efficacy to determine meaningful benefit and provide a basis for sample size estimation in clinical trials.

Adult↗

The fate of endothelial function testing: rationale and design of the Firefighters And Their Endothelium (FATE) study.

Endothelial dysfunction plays a pivotal role in the development and progression of atherosclerotic vascular disease. The endothelium is strategically located between blood and vascular smooth muscle, making it both vulnerable to a variety of injurious stimuli but also available for interrogation as a marker of vascular health. Firefighters And Their Endothelium (FATE) is a prospective, longitudinal cohort study designed to assess the relationship between endothelial function, emerging cardiovascular risk factors and ultimately atherosclerotic vascular disease. It is hypothesized that participants with impaired endothelial function will be at increased risk of atherosclerotic complications. This Canadian initiative will recruit 1600 middle-aged participants with no known history of cardiovascular disease to be followed for cardiovascular events over the next decade. Quantitative B-mode ultrasound will be employed to assess endothelial function and subclinical atherosclerosis. This research is designed to redefine the approach to the primary prevention of atherosclerosis.

Adult↗

Radioprotection by WR-151327 against the late normal tissue damage in mouse hind legs from gamma ray radiation.

PURPOSE: To evaluate the protective effect of WR-151327 on late radiation-induced damage to normal tissues in mice. METHODS AND MATERIALS: The right hind legs of mice with or without WR-151327 administration (400 mg/kg) were irradiated with 137Cs gamma rays. Leg contracture and skin shrinkage assays were performed at 380 days after irradiation. The mice were killed on day 400 postirradiation and histological sections of the legs were made. The thickness of the dermis, epidermis, and skin (dermis plus epidermis) was measured. The muscular area of the legs and the posterior knee angle between the femur and tibia were also measured. The left hind legs were similarly assessed as nonirradiated controls. Group means and standard deviations were calculated and dose-response curves were drawn for every endpoint. Then, the dose modifying factor (DMF) for each endpoint and the correlations among endpoints were determined. RESULTS: Late damage assayed by leg contracture and skin shrinkage progressed with increasing radiation dose. However, it was reduced by drug treatment. The significant effect was indicated for skin shrinkage by a DMF of 1.8 at 35%. The DMF for leg contracture was 1.3 at 6 mm. In the irradiated legs, epidermal hyperplasia and dermal fibrosis in the skin, muscular atrophy, and extension disturbance of the knee joint were observed. These changes progressed with increasing radiation dose. Skin damage assayed by the present endpoints was also reduced by drug treatment by DMFs of 1.4 to 1.7. However, DMFs for damage to the muscle and knee were not determined because no isoeffect was observed. There were good correlations between leg contracture or skin shrinkage and the other endpoints in both untreated and drug-treated mice. CONCLUSIONS: WR-151327 has the potential to protect against radiation-induced late normal tissue damage.

Animals↗