Impact of different methodologies on the detection of point mutations in routine air-dried fine needle aspiration (FNA) smears.

Currently the best method to select suspicious thyroid nodules for surgery is fine needle aspiration (FNA) cytology. However, FNA cytology has some inherent limitations, which can partly be overcome by molecular analysis. Therefore, molecular testing for somatic mutations has emerged as the most promising approach for molecular FNA diagnostics. The objective of this methodological study was to evaluate the feasibility of detecting BRAF, NRAS, HRAS, and KRAS mutations from routine air-dried thyroid FNA smears, and to find an optimal method for detecting these mutations in FNA samples. DNA was extracted from 110 routine air-dried FNA smears and the corresponding surgically obtained formalin-fixed paraffin-embedded tissues. The presence of BRAF, NRAS, HRAS, and KRAS mutations was assessed by real-time PCRs and high resolution melting analysis, and/or pyrosequencing in comparison to real-time PCRs using hybridization probes and fluorescence melting curve analysis. The high-resolution melting-PCRs revealed a significantly lower number of PCR failures and questionable results, and detected more mutations than the PCRs using hybridization probes. The number of PCR failures ranging from 14-16% by high-resolution melting-PCRs could be further reduced to 5-14% by adding pyrosequencing assays. Moreover, pyrosequencing increased the specificity of the assays, up to 98-100%, while the sensitivity ranged between 32-63%. In summary, the mutation detection, especially in air-dried FNA samples, improves when using PCR assays in combination with high resolution melting analysis. Additional improvement can be obtained by subsequent pyrosequencing in comparison to previously described real-time PCRs using hybridization probes and fluorescence melting curve analysis.

Oral health-related quality of life, and satisfaction with treatment and treatment outcomes of adolescents/young adults with cleft lip/palate: an exploration.

The objectives of this correlational study were to explore whether the oral health-related quality of life (ohrqol) of adolescents/young adults with cleft lip/palate (CL/P) relates to their own and their parents' satisfaction with treatment and treatment outcomes. Data were collected using mailed surveys from 30 parents and 27 patients who completed CL/P treatment. Patients' ohrqol was assessed with the Michigan Oral Health-Related Quality of Life (MOHRQoL) Scale, treatment satisfaction with Kiyak's Post-Surgical Patient Satisfaction Questionnaire, and satisfaction with treatment outcomes with Noor and Musa's Cleft Evaluation Profile. Overall, patients reported a positive ohrqol. Their satisfaction scores ranged from low to high. Patient and parent treatment satisfaction was related, while their outcome satisfaction did not correlate. Patients' MOHRQoL scores correlated with parent and patient treatment satisfaction and parents' outcome satisfaction. Patients' MOHRQoL scores did not correlate with patients' outcome satisfaction. The patients' level of discomfort was strongly correlated with patients' and parents' treatment satisfaction and parents' outcome satisfaction. In conclusion, regardless of outcome satisfaction, young patients with CL/P report a quite positive ohrqol after treatment completion. There is a clear relationship between the ohrqol of these patients with CL/P and their own as well as their parents' treatment satisfaction assessments.

Novel sperm-binding proteins of epididymal origin contain four fibronectin type II-modules.

Novel fibronectin type II (Fn2)-module proteins were cloned from human and canine epididymal cDNA libraries. cDNA sequences predicted a highly conserved protein family, related but not homologous to ungulate seminal plasma proteins (approximately 50% sequence identity), and the first known examples of proteins with four tandemly arranged Fn2-domains. By Northern blot and in situ hybridization analyses the encoding mRNAs were shown to be abundant products of the epididymal duct epithelium, but not detectable in other tissues. Homologous mRNAs were identified in the epididymides of various mammals, representing members of this novel protein family of epididymal origin. Within the Fn2-module-encoding stretches, species homologues displayed >85% sequence identity, but showed high variability at their predicted N-termini. An antipeptide antiserum in Western blot analyses detected 30-35 kDa immunoreactive protein bands in epididymal tissue, cauda epididymidal fluid, and sperm membrane protein preparations. The tandem arrangement of increasing numbers of Fn2-modules might functionally correspond to the tendency to form oligomers that has been described for lipid-binding proteins.

Incidence of hydroxychloroquine retinopathy in 1,207 patients in a large multicenter outpatient practice.

OBJECTIVE: To define the true risk of hydroxychloroquine (HCQ) retinal toxicity by studying the largest single group yet evaluated. METHODS: Retrospective chart review of all patients in the Kaiser Permanente Medical Care Program, Southern California Region, who had HCQ prescriptions filled from 1991 through 1993 (1,556 patients in 11 medical centers). Of 1,207 charts of patients who took HCQ and had documented ophthalmologic examinations, initial screening identified 21 charts (1.7%) that indicated possible HCQ toxicity. RESULTS: We identified 1 patient with definite toxicity (1 of 1,207; 0.08%) and 5 other patients with indeterminate but probable toxicity (5 of 1,207; 0.4%). The incidence of definite HCQ retinal toxicity in patients treated with HCQ at <6.5 mg/kg/day was 0. CONCLUSION: In HCQ-treated patients whose renal function is normal, routine ophthalmic screening is not indicated if the daily dosage is <6.5 mg/kg. In patients whose daily dosage is >6.5 mg/kg or who have taken HCQ continuously for > 10 years, annual screening may be appropriate.

A further test of the inverted-U hypothesis relating achievement anxiety and academic test performance.

The assumption that the inverted-U hypothesis, which shows performance as a function of activation level, mediates the relationship between achievement anxiety and academic test performance was tested by comparing Achievement Anxiety Test scores of 75 male and female college students with a self-report measure of activation taken prior to a classroom examination. Results supported the predicted relationship between achievement anxiety reaction type and academic performance (rho less than .05), but only partially supported the inverted-U hypothesis posited to account for this relationship. Results were further interpreted as suggesting that examinees experience two general types of arousal in the testing situationone type that enhances performance and one that impedes performance. Further implications of the results were discussed.