Cloning and expression of surface antigens from occult chronic hepatitis B virus infections and their recognition by commercial detection assays

Occult hepatitis B virus (HBV) infections show little or no serological markers of viral infection, including the absence of hepatitis B surface antigen (HBsAg) which is the main marker of ongoing HBV infection. Such infections can be important in the context of blood and/or organ donations. To study whether mutations contribute to HBsAg seronegativity, S gene sequences from such patients were amplified and cloned. Sequencing revealed 12 clones from seven different patients which contained potentially important mutations. The sequences were subcloned into an expression vector and mutant HBsAgs were expressed in cell culture. The capacity of three HBsAg detection assays to recognise the mutant HBsAgs was studied. Three categories were found: mutant HBsAgs that are not recognised by the assays, those that are recognised as well as wild-type (WT) antigen and an intermediate category where detection of the mutant HBsAgs is reduced with respect to WT. Most of the isolates fall into the second category. Mutations can therefore contribute to HBsAg seronegativity in occult HBV infections, but in most cases the explanation is probably the low level of viral replication.     

Disease prevention and health promotion in medical education: reflections from an academic health center.

PURPOSE: It is unclear whether academic health centers are successfully addressing societal needs and expectations by preparing students with knowledge and skills in disease prevention and health promotion. The authors assessed whether students were exposed to key content in these areas and whether they felt this exposure was adequate. METHOD: All components of the first three years of the Case Western Reserve University (Case) curriculum were examined in 2001 to create a curricular map, using competencies in disease prevention and health promotion identified by the Association of Teachers of Preventive Medicine (ATPM) as a template to assess the scope of instruction. Case students' United States Medical Licensing Examination (USMLE) Step 2 subscores in preventive medicine and health maintenance from 1994 to 2000 and graduating seniors' assessment of the adequacy of their training were compared to national data from the Association of American Medical Colleges' 2000 Graduation Questionnaire (GQ). RESULTS: Most content areas identified by ATPM were present in the Case curriculum and were offered frequently in a variety of educational venues over the first three years. USMLE scores increased nationally and at Case from 1994 to 2000 and Case students' perception of training adequacy in preventive medicine and health promotion was comparable to national ratings from the 2000 GQ. CONCLUSIONS: Broad and frequent exposure to disease prevention and health promotion core competencies has value, but may not sufficiently prepare students to deliver health-promoting services confidently. Creative curricula highlighting prevention's relevance throughout clinical practice and incorporating formal opportunities to apply knowledge and build experience may result in greater success.     

Elucidation of ataxin-3 and ataxin-7 function by integrative bioinformatics

The spinocerebellar ataxias (SCAs) are a class of hereditary neurodegenerative diseases, which are caused by the pathological expansion of unstable CAG triplet repeats found in a number of apparently unrelated genes. The proteins encoded by the SCA genes typically translate this expanded (CAG)n repeat into an expanded poly(Q) stretch. Several pathological features are common to all SCAs, irrespective of the gene harbouring the expansion. The specific contributions of the mutated genes are currently hard to assess, as the physiological role of most of the so-called ataxins is not known. By combining the results of profile-based sequence analysis with genome-wide functional data available for model organisms, we have derived detailed predictions of the physiological function of two SCA gene products. Ataxin-3, the protein mutated in Machado Joseph Disease (SCA3), belongs to a novel group of cysteine-proteases and is predicted to be active against ubiquitin chains or related substrates. The catalytic site of this enzyme class is similar to that found in UBP and UCH type ubiquitin proteases. For ataxin-7, the gene product of the SCA7 gene, we have identified an orthology relationship to the yeast open reading frame Ygl066c. Recently published evidence from genome-wide studies suggests that Ygl066c is a component of the SAGA histone acetyltransferase complex. By analogy, a similar role for the mammalian ataxin-7 can be expected. The functional predictions reported here are sufficiently precise to allow a direct experimental verification. Moreover, both findings have implications for the general pathogenesis of spinocerebellar ataxias by providing a direct connection of these diseases with ubiquitin metabolism and histone acetylation.     

National Department of Defense surveillance data for antibiotic resistance and emm gene types of clinical group A streptococcal isolates from eight basic training military sites

Antibiotic resistance and emm gene types were examined from 692 Group A streptococci isolates from eight United States military basic training sites between 1998 and 2001. Macrolide resistance was associated with geographic sites and emm type. These data are useful for vaccine development initiatives and antimicrobial treatment considerations.     

Effects of nitric oxide synthase inhibition by l-NAME on oxygen uptake kinetics in isolated canine muscle in situ

We hypothesized that an acute bout of strenuous, non-damaging exercise would increase rates of protein synthesis of collagen in tendon and skeletal muscle but these would be less than those of muscle myofibrillar and sarcoplasmic proteins. Two groups ( n = 8 and 6) of healthy young men were studied over 72 h after 1 h of one-legged kicking exercise at 67% of maximum workload ( W _max). To label tissue proteins in muscle and tendon primed, constant infusions of [1-¹³C]leucine or [1-¹³C]valine and flooding doses of [¹⁵N] or [¹³C]proline were given intravenously, with estimation of labelling in target proteins by gas chromatography–mass spectrometry. Patellar tendon and quadriceps biopsies were taken in exercised and rested legs at 6, 24, 42 or 48 and 72 h after exercise. The fractional synthetic rates of all proteins were elevated at 6 h and rose rapidly to peak at 24 h post exercise (tendon collagen (0.077% h⁻¹), muscle collagen (0.054% h⁻¹), myofibrillar protein (0.121% h⁻¹), and sarcoplasmic protein (0.134% h⁻¹)). The rates decreased toward basal values by 72 h although rates of tendon collagen and myofibrillar protein synthesis remained elevated. There was no tissue damage of muscle visible on histological evaluation. Neither tissue microdialysate nor serum concentrations of IGF-I and IGF binding proteins (IGFBP-3 and IGFBP-4) or procollagen type I N-terminal propeptide changed from resting values. Thus, there is a rapid increase in collagen synthesis after strenuous exercise in human tendon and muscle. The similar time course of changes of protein synthetic rates in different cell types supports the idea of coordinated musculotendinous adaptation.     

Therapist affirmation and the process and outcome of psychotherapy: two sequential analytic studies

Therapist affirmation was examined in two studies, the first with cognitive-behavioral therapy (CBT, N = 22) and the second with time-limited dynamic psychotherapy (TLDP, N = 20). Affirmation was identified using Structural Analysis of Social Behavior (SASB), and sequential analysis used to gauge its impact on therapy process and outcome. It was hypothesized that affirmation would correspond with patients' continuation of the current topic of conversation, but relate negatively to outcome if generally offered after maladaptive statements. Patients in Study 1 (CBT) were more likely to continue a topic after it was affirmed, and affirmation of maladaptive statements reliably differentiated patients who evidenced clinically significant change from those who did not at termination and at 12-months posttreatment. Affirmation of maladaptive statements in Study 2 (TLDP) corresponded with poorer outcome in relationships with significant others, but unexpectedly with improvements in self-concept.     

Exclusion of the alpha 1(II) cartilage collagen gene as the mutant locus in type IA osteogenesis imperfecta.

Using two restriction site polymorphisms within the structural gene coding for human type II collagen we have examined the segregation of this gene in three pedigrees with dominantly inherited osteogenesis imperfecta (Sillence type IA). We have demonstrated that the gene does not segregate with clinical expression of the disease and cannot, therefore, contain the mutation responsible for osteogenesis imperfecta in these families.     

Hostility and sex differences in the magnitude, duration, and determinants of heart rate response to forehead cold pressor: Parasympathetic aspects of risk

Recent models hypothesize that hostility confers increased risk of CHD through weaker parasympathetic dampening of cardiovascular reactivity (CVR). We tested this possibility using the forehead cold pressor task, a common maneuver which elicits the “dive reflex” characterized by a reflexive decrease in HR presumably through cardiac-parasympathetic stimulation. Participants were initially chosen from the outer quartiles of a sample of 670 undergraduates screened using the hostility subscale of the Aggression Questionnaire ([Buss, A.H., Perry, M., 1992. The Aggression Questionnaire. Journal of Personality and Social Psychology, 63, 452-459.]). The final sample of 80 participants was evenly divided between men and women and high and low hostility. Following a 10-min baseline, participants underwent a 3-min forehead cold pressor task. The task evoked a significant HR deceleration that was mediated by PNS activation, as assessed by respiratory sinus arrhythmia (RSA). Replicating prior research, men displayed greater decrease in HR. More important, low hostiles maintained larger HR deceleration over time compared to high hostiles although the autonomic basis for this effect was unclear. The findings broaden understanding of hostility and sex-related cardiovascular functioning and support the task as a method for evoking PNS-cardiac stimulation.