Evaluation of the recombinant 38-kilodalton antigen of Mycobacterium tuberculosis as a potential immunodiagnostic reagent

The diagnosis of infection caused by Mycobacterium tuberculosis is of increased public health concern following increases in the number of cases in developed countries and major increases in developing countries associated with the spread of human immunodeficiency virus (HIV) infection. The specificity of purified protein derivative skin testing for the detection of infection is compromised by exposure to environmental mycobacteria. Examination of sputum detects the most infectious patients, but not those with extrapulmonary disease. The 38-kDa antigen of M. tuberculosis contains two M. tuberculosis-specific B-cell epitopes. We overexpressed the gene for this antigen in Escherichia coli and evaluated the recombinant product in in vitro assays of T-cell function and as a target for the antibody response in humans. The sensitivity and specificity of the antigen as a skin test reagent were also assessed in outbred guinea pigs. We found that 69% of healthy sensitized humans recognize the antigen in vitro, as manifested by both cell proliferation and the production of gamma interferon. Untreated patients initially have a lower frequency of response (38%); this recovers to 72% during therapy. A total of 292 patients (20 with HIV coinfection) and 58 controls were examined for production of antibody to the 38-kDa antigen by using a commercially available kit. The sensitivity of the test in comparison with that of culture was 72.6%, and the specificity was 94.9%. The antigen was also tested for its ability to induce skin reactions in outbred guinea pigs sensitized by various mycobacterial species. The antigen provoked significant skin reactions in M. tuberculosis-, M. bovis BCG-, and M. intracellulare-sensitized animals. The significance of these findings and the usefulness of this antigen in immunodiagnosis are discussed.     

Excessive scapular motion in individuals recovering from painful and stiff shoulders: causes and treatment strategies

BACKGROUND AND PURPOSE: Scapular excursion and the wrist speed were studied before and after instruction and practice designed to achieve symmetrical scapular movement. SUBJECTS: Subjects were 10 female and 6 male patients, aged 44 to 78 years (chi=60.3, SD=11.2), with diagnosed shoulder pathologies. METHODS: Subjects were videotaped performing a reaching task. Pain status was monitored. The subjects were instructed to make the scapular movement symmetrical. They then repeated the task, while being videotaped, to monitor the effect of instruction. RESULTS: Individuals with asymmetric upper-extremity starting positions had excessive vertical motion of the involved scapula, which they controlled after instruction. Peak wrist speed of the involved upper extremity was lower only after instruction. CONCLUSION AND DISCUSSION: Even in the absence of biomechanical factors or pain, excessive scapular vertical motion appears to occur in the involved upper extremities of individuals recovering from unilateral shoulder problems. Improved scapular control can follow simple verbal instruction and practice, with a slight decrement in wrist speed.     

Structural studies of the putative O-specific polysaccharide of Acinetobacter baumannii O2 containing 3,6-dideoxy-3-N-(D-3-hydroxybutyryl)amino-D-galactose

A polysaccharide containing D-galactose, 2-deoxy-2-N-acetylamino-D-galactose and 3,6-dideoxy-3-N-(D-3-hydroxybutyryl)amino-D-galactose, probably corresponding to the lipopolysaccharide side chain, was obtained from an aqueous phenol extract of isolated cell walls from Acinetobacter baumannii strain O2. By means of NMR studies and chemical degradations, the repeating unit of the polymer was identified as a branched hexasaccharide of the structure shown, where Fuc3N represents 3-amino-3,6-dideoxygalactose and R represents D-3-hydroxybutyryl. Serological tests indicated that the polymer corresponded to the O2 antigen.     

The prevalence of clinically significant anaemia and haemoglobinopathy in children requiring dental treatment under general anaesthesia: a retrospective study of 1000 patients

There is little information on the possible value of screening children who are liable to haemoglobinopathies as part of pre-operative assessment for dental treatment under general anaesthesia. The present retrospective investigation examined, first, the number of patients having low haemoglobin levels among 1000 patients who had undergone haematological investigation prior to general anaesthesia in a dental outpatient unit, and, secondly, the subsequent clinical management of these patients. Haemoglobin levels of 10.0 g/dl or less were found in 31 children: 13 Asian, 7 Afro-Caribbean, 5 Mediterranean, 3 Arabic, 1 white Caucasian and 2 Oriental children. In addition, 17 patients had sickle-cell trait and 2 had beta-thalassaemia trait, but there was no relationship between the presence of haemoglobinopathy and low levels of haemoglobin. The planned general anaesthesia was undertaken for 22 of the 31 children who had low levels of haemoglobin and for the 19 children with haemoglobinopathy. Only 6 children ultimately did not undergo general anaesthesia, all failing to return. It is concluded that pre-anaesthetic haematological assessment of children needing minor dental surgery is rarely of any significant clinical value.     

Cisapride reduces neonatal postoperative ileus: randomised placebo controlled trial

AIM: To assess the efficacy of cisapride in reducing ileus persisting to the tenth postoperative day after neonatal abdominal surgery. METHODS: A prospective, randomised, double blind trial comparing rectal cisapride (1.4-2.3 mg/kg/day) with placebo over seven days was undertaken in 33 neonates. RESULTS: Seven of 12 (58%) patients receiving placebo and eight of 11 (73%) receiving cisapride achieved a first sustained feed during treatment. Of those receiving cisapride, the first sustained feed occurred at 2.3 days (SEM 0.6) compared with 4.7 days (SEM 0.8) with placebo. By the seventh day the mean daily net enteral balance was 69 (SEM 18) ml/kg in the cisapride subgroup and 17 (SEM 8) ml/kg for those receiving placebo. Stool was passed on 6.3 (SEM 0.4) treatment days in the cisapride subgroup compared with 4.1 (SEM 1.0) treatment days in the placebo subgroup. CONCLUSION: Cisapride is effective in neonates with a prolonged ileus after abdominal surgery.     

SED88: a Pascal program for the analysis of sedimentation equilibrium data

Analytical ultracentrifugation is commonly used for the determination of molecular weights (sedimentation equilibrium) and sedimentation coefficients (sedimentation rate) of biological macromolecules in solution. A Turbo Pascal program for the analysis of sedimentation equilibrium centrifugation data produced by absorbance optical systems is described. The user may enter data from a scan of absorbance versus distance from the centre of rotation, via a graphics tablet (or ASCII file). This is subsequently manipulated to yield an apparent weight average molecular weight for the given sample. Plots of ln (absorbance) versus (radius2) may also be produced. The method described uses readily available computational equipment requiring only a graphics tablet in addition to an IBM PC compatible computer. This technique and the software developed have been used to investigate the molecular weight range of two International Humic Substances Society (IHSS) reference samples from the Suwannee River.     

Calcium phosphate/calcium oxalate crystal association in urinary stones: implications for heterogeneous nucleation of calcium oxalate

PURPOSE: Most stones contain more than one type of crystals, and some combinations, such as calcium phosphate/calcium oxalate, are more common than others. Epitaxy between the crystals has been suggested to play a role in growth of such stones. The specific aim of this study is to investigate the involvement of calcium phosphate in crystallization of calcium oxalate. MATERIALS AND METHODS: Twenty calcium oxalate stones or stone fragments were examined using various microscopic techniques, including scanning, transmission and back-scattered electron microscopy. Similarly, calcium oxalate stones induced on a plastic foreign body implanted inside urinary bladders of laboratory rats were also investigated. Examination of the interface between calcium phosphate and calcium oxalate crystals was emphasized. RESULTS: Close association between crystals of calcium phosphate and calcium oxalate were found in both the human and rat stones. All crystals examined were associated with an organic matrix on the surface and contained copious amounts of organic material within the crystalline entities. Interface between the crystals also appeared to be occupied by organic matrix. CONCLUSIONS: Results of this and other studies from our laboratory indicate that epitaxy between various crystals, even though theoretically possible, appears unlikely in vivo. The appearance of specific crystalline combinations in stones is probably a result of the urinary environment being conducive for crystallization of those components. Heterogeneous nucleation of calcium oxalate is most probably induced by biological elements, including membranous cellular degradation products.     

Homologous and heterologous desensitisation of somatostatin-induced increases in intracellular Ca2+ and inositol 1,4,5-trisphosphate in CHO-K1 cells expressing human recombinant somatostatin sst5 receptors

The mechanisms responsible for somatostatin (SRIF)-induced increases in intracellular Ca2+ concentration ([Ca2+]i) and subsequent desensitisation were studied in CHO-K1 cells expressing human sst5 receptors (CHOsst5 cells). To study the nature of the desensitisation, interactions with uridine triphosphate (UTP) were examined. SRIF (pEC50 7.10) and UTP (pEC50) 5.14) caused concentration-dependent increases in [Ca2+]i but the SRIF maximum was about 40% of that to UTP. SRIF-, but not UTP-, induced increases in [Ca2+]i were transient and abolished by pertussis toxin. SRIF and UTP caused sustained increases in Ins(1,4,5)P3 but the SRIF maximum was about 30% of that to UTP. Removal of [Ca2+]e attenuated the SRIF-induced peak rise in [Ca2+]i but had no effect on the peak increases in Ins(1,4,5)P3. UTP-induced increases in [Ca2+]i and Ins(1,4,5)P3 were attenuated in the absence of [Ca2+]e. Following pre-exposure to SRIF (1 microM) or UTP (100 microM) for 5 min, subsequent SRIF responses were desensitised. Similar results were obtained in the absence of [Ca2+]e. Pre-exposure to SRIF had no effect on subsequent responses to UTP but in the absence of [Ca2+]e, responses to UTP were attenuated. The results suggest that SRIF but not UTP-induced increases in [Ca2+]i in CHOsst5 cells are mediated by pertussis toxin sensitive G proteins and are caused by an entry of extracellular Ca2+ and release from an Ins(1,4,5)P3 sensitive Ca2+ store. Homologous or heterologous desensitisation of agonist-induced increases in [Ca2+]i could be demonstrated in the presence or absence of extracellular Ca2+ respectively, and the latter appeared to involve depletion of a common intracellular Ca2+ store.