Ordered shotgun sequencing of a 135 kb Xq25 YAC containing ANT2 and four possible genes, including three confirmed by EST matches

Ordered shotgun sequencing (OSS) has been successfully carried out with an Xq25 YAC substrate. yWXD703 DNA was subcloned into lambda phage and sequences of insert ends of the lambda subclones were used to generate a map to select a minimum tiling path of clones to be completely sequenced. The sequence of 135 038 nt contains the entire ANT2 cDNA as well as four other candidates suggested by computer-assisted analyses. One of the putative genes is homologous to a gene implicated in Graves' disease and it, ANT2 and two others are confirmed by EST matches. The results suggest that OSS can be applied to YACs in accord with earlier simulations and further indicate that the sequence of the YAC accurately reflects the sequence of uncloned human DNA.     

In vitro isolation and identification of human immunodeficiency virus (HIV) variants with reduced sensitivity to C-2 symmetrical inhibitors of HIV type 1 protease

Protease inhibitors are another class of compounds for treatment of human immunodeficiency virus (HIV)-caused disease. The emergence of resistance to the current anti-HIV drugs makes the determination of potential resistance to protease inhibitors imperative. Here we describe the isolation of an HIV type 1 (HIV-1) resistant to an HIV-protease inhibitor. Serial passage of HIV-1 (strain RF) in the presence of the inhibitor, [2-pyridylacetylisoleucylphenylalanyl-psi (CHOH)]2 (P9941), failed to yield a stock of virus with a resistance phenotype. However, variants of the virus with 6- to 8-fold reduced sensitivity to P9941 were selected by using a combination of plaque assay and endpoint titration. Genetic analysis and computer modeling of the variant proteases revealed a single change in the codon for amino acid 82 (Val-->Ala), which resulted in a protease with lower affinity and reduced sensitivity to this inhibitor and certain, but not all, related inhibitors.     

Exercise testing and prescription. Practical recommendations for the sedentary

Past research on help-seeking behaviour and compliance among the mentally ill has mainly been focused on the examination of individual and structural determinants, while the impact of the socio-cultural context has been largely neglected. To examine this impact, we conducted a representative survey among German citizens who were eligible to vote. By means of vignettes describing different mental disorders we recorded the lay public's preferences for various treatment methods, together with their subjective reasons for voicing these particular preferences. While we found that psychotherapy was generally held in high esteem by the lay public, psychopharmacotherapy was rejected by the vast majority of respondents. This pattern was observed for all of the mental disorders included in our survey. While the public's image of psychotherapy is largely determined by popular views on psychoanalysis, public opinion about psychotropic drugs is strongly influenced by characteristics associated with tranquillizers. Among the different psychotherapeutic approaches, psychoanalysis is the preferred method among respondents in the western part of Germany, while the lay public in the eastern part tends to endorse group therapy. Finally, our research showed a considerable difference between the conceptions held by the lay public and those of psychiatric experts with regard to the adequate treatment of mental disorders. This, in turn, should not be without consequence for help-seeking behaviour and patient compliance.     

Recombinant tumor necrosis factor for superficial bladder tumors

Twenty patients with histologically documented superficial bladder cancer (Ta, T1, Tis) were treated with intravesical administration of TNF 400-1800 micrograms. Of 18 patients with a marker lesion, 2 obtained a complete response for 8+ and 18 months. Two had a partial response and were given other intravesical therapies after 5 and 7 months. No or minimal systemic absorption of TNF was observed and documented in 4 of 20 patients by pharmacokinetic studies, and no patients developed antibodies to intravesically administered TNF. TNF was well tolerated in doses up to 1800 micrograms. No systemic or local side effects were observed. Modest activity was attained with intravesical TNF, even in pretreated patients.     

Investigation of a synthetic peptide as immunogen for a variant epidermal growth factor receptor associated with gliomas

We have previously demonstrated antibody production to a glioma-associated variant form of the human epidermal growth factor receptor in rabbits that had received a synthetic peptide mimicking the unique primary structure of the variant protein as immunogen. We report here the response of mice, rabbits, goats, and macaques immunized by various protocols to this peptide. Titers to both peptide- and cell-elaborated variant receptor were measured, and the capacity to recognize the variant receptor in human tumor samples was determined. Within the range of species and strains investigated, we demonstrated a variable species-associated response to the peptide (rabbits > mice > goats > rats > macaques). Rabbits and a single goat produced specific, high titer antibody activity to the variant receptor protein following immunization with peptide alone. Murine titers to the parent protein were not appreciable following peptide immunization alone; additional immunization with variant receptor as expressed on cell membranes was used to boost this response.     

Epidemiology of acute hepatitis B in a university hospital in S?o Paulo, Brazil: retrospective study of two five-year periods

CONTEXT: HBV infection is endemic in Brazil and acute HBV infection is still a common disease. OBJECTIVE: To analyze incidence, risk factors and evolution of acute HBV infection. SETTING: University Hospital. PATIENTS: 357 patients with acute HBV infection, comparing two periods: 1985-1989 vs. 1990-1994. RESULTS: The overall incidence declined from 50 new cases/year in 1985-89 (30% of all cases) to 25 new cases/year in 1990-94 (8% of all cases). Transmission among male homosexuals (3.9% of cases in 85-89) declined to 1.3% in 90-94 (p > 0.05). Amongst health care workers (HCW) it declined from 8.2% to 2.0% (p = 0.02). Conversely, heterosexual transmission increased from 4.8% to 10.1% (p = 0.06). Chronification of HBV infection following the acute episode was observed in 1.7%. Fulminant hepatic failure was seen in 3.4%. However, 27.2% of patients were lost to follow-up before normalization of the biochemical tests. CONCLUSIONS: The different patterns of risk factors observed is probably related to measures for preventing AIDS and to HCW vaccination programs. Chronification following acute episodes was not a common event.     

The perils of permanent punctal plugs

Satisfactory pain relief with postoperative extradural bupivacaine increases the amount of breast feeding after Caesarean section. To investigate the effect of extradural buprenorphine, we have evaluated the amount of breast feeding and the gain in infant weight for 11 days after Caesarean section in patients who received continuous extradural bupivacaine with or without buprenorphine. Extradural buprenorphine significantly decreased both measures although there was no significant difference in pain intensity. We suggest that extradural buprenorphine suppressed breast feeding after Caesarean section.     

Bactericidal action of ofloxacin, sulbactam-ampicillin, rifampin, and isoniazid on logarithmic- and stationary-phase cultures of Mycobacterium tuberculosis

The bactericidal actions of ofloxacin and sulbactam-ampicillin, alone and in combination with rifampin and isoniazid, on exponential-phase and stationary-phase cultures of a drug-susceptible isolate of Mycobacterium tuberculosis were studied in vitro. In exponential-phase cultures, all drugs were bactericidal, with the higher concentrations of ofloxacin (5 micrograms/ml) and sulbactam-ampicillin (15 micrograms of ampicillin per ml) being as bactericidal as 1 microgram of isoniazid per ml or 1 microgram of rifampin per ml. In two-drug combinations, both drugs increased the levels of activity of isoniazid and rifampin and were almost as bactericidal as isoniazid-rifampin; they also appeared to increase the level of activity of isoniazid-rifampin in three-drug combinations. In contrast, ofloxacin and sulbactam-ampicillin had little bactericidal activity against stationary-phase cultures and were less active than isoniazid or rifampin alone. Furthermore, in two-drug or three-drug combinations, they did not increase the level of activity of isoniazid, rifampin, or isoniazid-rifampin. These findings suggest that ofloxacin and sulbactam-ampicillin are likely to be most useful in the early stages of treatment and in preventing the emergence of resistance to other drugs but are unlikely to be effective as sterilizing drugs helping to kill persisting lesional bacilli.