Rapid diagnosis of pneumonia in children

Antigen detection techniques are available for the identification of bacterial polysaccharides, viruses, and chlamydia. Viruses and chlamydia are detected by direct immunofluorescence (DFA) or enzyme immunoassay (EIA). Bacterial polysaccharides are detected by latex agglutination or staphylococcal coagglutination of serum or concentrated urine. Most studies have not compared these techniques to the gold standard of lung puncture, so the role of dual infections with bacteria and viruses cannot be adequately determined. The sensitivity of any of these techniques is dependent on the quality of the antisera used. Monoclonal sera are now available for the detection of most viruses and seem to be as sensitive as polyclonal sera. DFA or EIA may offer equal sensitivity but their advantages and disadvantages must be considered by the local diagnostic laboratories. Most DFA and EIA systems have a sensitivity of 90% when compared with viral cultural for the identification of the organism. Agglutination reagents are available commercially for the detection of pneumococcal and Hemophilus influenzae type b polysaccharides. The sensitivity and specificity of each brand should be determined on serum or urine from patients known to have positive blood cultures and those free of disease. The brand chosen should be the one that has reasonable sensitivity and specificity. Rapid diagnostic techniques are helpful if they are used within a clinical context and they are positive. Negative tests do not rule out infection.     

Intra-operative support via augmented reality techniques in endoscopic surgery.

Computer assisted operation planning systems are gaining increasing recognition in the field of surgery. These systems offer new possibilities for preparing an intervention, with the goal of reducing the amount of expensive operating-room time required for the intervention. The safest and most effective surgical approach should always be selected, but it is often difficult to transfer the output of the planning system to the intra-operative situation so that the planning results can be considered during the actual intervention. At the Fraunhofer Institute for Computer Graphics (IGD) in Darmstadt and the Centre for Advanced Media Technology (CAMTech) in Singapore methods are being developed to bridge the gap between the external planning session and the intra-operative case: Augmented Reality (AR) techniques are used to overlay preoperative scanned image data, as well as results of the planning session, on the operation field.     

Prolongation of allograft and xenograft survival in mice by anti-CD2 monoclonal antibodies.

Anti-CD2 monoclonal antibodies (mAb) were used to influence graft survival in two transplantation models. Xenogeneic rat islets were transplanted intraportally into mice. Anti-CD2 mAb prolonged xenograft survival and was synergistic with UVB irradiation in prolonging survival. Anti-CD2 mAb was also more potent than an anti-CD4 mAb in this model. Allogeneic cardiac grafts were transplanted across an entire H-2 difference and anti-CD2 mAb prolonged allograft survival in a dose-dependent fashion. Kinetic experiments revealed that anti-CD2 mAb was most potent when administered at the time of allografting. A delay in administration of mAb markedly reduced its immunosuppressive effects. Furthermore, additional doses of mAb given after the initial doses provided no increased immunosuppression and anti-CD2 mAbs did not delay rejection of second-set allografts. These findings support the notion that anti-CD2 mAbs interfere with afferent immunity and that CD2 is most important during the initial steps of an immune response. Investigation of the effect of anti-CD2 mAb on cellular immune functions demonstrated, in agreement with previous results, that it caused antigenic down-modulation of CD2 with relative sparing of CD3, CD4, and CD8 cell surface expression. Concomitantly the MLR, CTL, and NK responses were suppressed.     

Cytomegalovirus Disease in High-Risk Transplant Recipients Despite Ganciclovir or Valganciclovir Prophylaxis

The clinical patterns and predictors of cytomegalovirus (CMV) disease in kidney and/or pancreas transplant patients on ganciclovir (1.0 g po t.i.d.) or valganciclovir (450 mg po q.d.) prophylaxis were studied. This is a retrospective analysis of 129 transplant recipients. Median follow up was 12 months (range, 6-18 months). The overall incidence of CMV disease at 1-year post-transplant was 14% (4% tissue-invasive, 10% noninvasive). Seventeen of 18 patients were diagnosed with CMV after completion of 3 months' prophylaxis (median 8 weeks, range, 2-28 weeks). Induction treatment with thymoglobulin, and Donor +/Recipient - CMV status were the strongest predictors for the development of CMV disease. Cytomegalovirus incidence was not different between patients treated with ganciclovir or valganciclovir (15 vs. 17%, respectively). Valganciclovir (450 mg q.d.) is as effective as oral ganciclovir in CMV prophylaxis. High-risk individuals might require higher doses or longer duration of valganciclovir treatment.     

Evaluation of prophylaxis against hepatitis B in a large municipal hospital.

BACKGROUND: Perinatal transmission of hepatitis B can be interrupted by the administration of hepatitis B vaccine and hepatitis B globulin to the infants of carrier mothers. Universal screening of pregnant women makes this strategy possible. METHODS: To evaluate the implementation of universal hepatitis B surface antigen screening of women giving birth at Kings County Hospital Center during 1988, we reviewed laboratory records to find all women with a positive test result who might give birth. We also randomly reviewed records of women who gave birth to live infants to determine the percentage of screening in the population. Infants' charts were reviewed for documentation of maternal hepatitis B surface antigen status and administration of hepatitis B immune globulin and vaccine. RESULTS: Sixty infants who lived long enough to receive antihepatitis B prophylaxis were distinguished out of a total of 5146 births. Screening was done for from 66.8% to 80.4% (95% confidence interval) of the mothers of these infants. Although 44 of 60 infants received hepatitis B immune globulin and 39 of 60 infants received vaccine, only 27 of 60 received vaccine within 12 hours in combination with immune globulin (Centers for Disease Control-recommended therapy). CONCLUSIONS: Documentation of hepatitis B surface antigen in the infant's delivery room record was present in 23 of 60 infants. Those infants all received hepatitis B immune globulin and vaccine; 21 received hepatitis B immune globulin within 12 hours. Hepatitis B immune globulin was given within 12 hours to 8 of 37 infants who lacked documentation of hepatitis B surface antigen status on the delivery room record. These differences were highly significant (p less than 0.001) even when only the 40 patients who had documented prenatal screening at Kings County Hospital Center (21/23 vs 4/17). Prenatal care did not have any effect on outcome.     

Development of an ELISA for detection of an organophosphorus compound using monoclonal antibodies

This paper describes a specific and highly sensitive ELISA system using monoclonal antibodies in order to assay an organophosphorus compound. The soman derivative methyl phosphonic acid, p-aminophenyl 1,2,2,-trimethyl-propyl diester (MATP) served as model substance. In order to obtain antibody-producing hybridomas BALB/c mice were immunized with MATP linked onto human serum albumin (HSA). The spleen cells of immunized mice were fused with syngenic plasmacytomas of the non-producer-line X63Ag8.653 with the aid of polyethylene glycol. To eliminate undesirable cross-reaction, common screening procedures were modified by directly coating the ELISA plates with hapten. Five out of 15 positive cell-lines were cloned by limiting dilution and further propagated. The respective immunoglobulin class and subclass of the obtained monoclonal antibodies was determined. Four of which were identified as IgG₁, the other as IgG_2a. After enrichment of antibodies in ascites and their isolation by protein A-sepharose, the affinity of various monoclonal antibodies was estimated in competitive inhibition enzyme immunoassay (CIEIA) by measuring the IC₅₀ rates of free MATP. The rates were found to lie between 2.5 × 10^–6 mol/l and 4.3 × 10^–4 mol/l MATP. The IC₁₀ rate for detectable MATP concentration was 5.4 × 10^–7 mol/l MATP. Test duration was 280 min. The reactivity of the monoclonal antibodies with structurally related substances was used to check their specificity. Cross-reaction turned out to be negative. In order to develop a direct competitive ELISA, MATP was linked to horse radish peroxidase (HRPO) by adding a spacer. This helped to reduce total duration to 40 min. The detection level was further reduced to 1.3 × 10^–7 mol/l MATP (corresponding to 975 pg/25 l test-buffer) using the monoclonal antibody F71D7. Likewise, MATP was detected in goat serum, chicken serum, rabbit serum, milk and company's water in concentrations between 2.1 × 10^–7 mol/l (IC₁₀, company's water) and 4.9 × 10^–8 mol/l (IC₁₀, milk).     

Committing embryonic stem cells to early endocrine pancreas in vitro

A panel of genetic markers was used to assess the in vitro commitment of murine embryonic stem (ES) cells toward the endoderm-derived pancreas and to distinguish insulin-expressing cells of this lineage from other lineages such as neuron, liver, and yolk sac. There are two nonallelic insulin genes in mice. Neuronal cells express only insulin II, whereas the pancreas expresses both insulin I and II. Yolk sac and fetal liver express predominately insulin II, small amounts of insulin I, and no glucagon. We found that ES-derived embryoid bodies cultured in the presence of stage-specific concentrations of monothio-glycerol and 15% fetal calf serum, followed by serum-free conditions, give rise to a population that expresses insulin I, insulin II, pdx-1 (a pancreas marker), and Sox17 (an endoderm marker). Immunohistochemical staining shows intracellular insulin particles, and its de novo production was confirmed by staining for C-peptide. Most, but not all, of the insulin+ or C-peptide+ cells coexpress glucagon, demonstrating a differentiation pathway to pancreas rather than yolk sac or fetal liver. Addition of beta-cell specification and differentiation factors activin beta B, nicotinamide, and exendin-4 to later-stage culture increased insulin-positive cells to 2.73% of the total population, compared with the control culture, which gave rise to less than 1% insulin-staining cells. These findings suggest that stepwise culture manipulations can direct ES cells to become early endocrine pancreas.