Cryptococcus neoformans capsule structure evolution in vitro and during murine infection

Cryptococcus neoformans capsule structure modifications after prolonged in vitro growth or in vivo passaging have been reported previously. However, nothing is known about the dynamics of these modifications or about their environmental specificities. In this study, capsule structure modifications after mouse passaging and prolonged in vitro culturing were analyzed by flow cytometry using the glucuronoxylomannan-specific monoclonal antibody E1. The capsule structures of strains recovered after 0, 1, 8, and 35 days were compared by using the level of E1-specific epitope expression and its cell-to-cell heterogeneity within a given cell population. In vitro, according to these parameters, the diversity of the strains was higher on day 35 than it was initially, suggesting the absence of selection during in vitro culturing. In contrast, the diversity of the strains recovered from the brain tended to decrease over time, suggesting that selection of more adapted strains had occurred. The strains recovered on day 35 from the spleen and the lungs had different phenotypes than the strains isolated from the brain of the same mouse on the same day, thus strongly suggesting that there is organ specificity for C. neoformans strain selection. Fingerprinting of the strains recovered in vitro and in vivo over time confirmed that genotypes evolved very differently in vitro and in vivo, depending on the environment. Overall, our results suggest that organ-specific selection can occur during cryptococcosis.     

Human platelets after their contact with influenza virus activate the complement system in autologous serum

Guinea pig erythrocytes that have been exposed to influenza virus activate the alternative pathway through virus-induced desialation of the cells. Neuraminidase treatment of rabbit platelets enhance their clearancein vivo. Washed human platelets were labeled with⁵¹Cr exposed to Influenza virus, and resuspended in autologous serum that had been dialyzed against Veronal-buffered saline containing Ca⁺⁺ and Mg⁺⁺ (VBS⁺⁺), VBS containing 8 mM EGTA and 2 mM Mg⁺⁺ (VBS-MgEGTA) or VBS containing 20 mM EDTA (VBS-EDTA) for 60 min at 37°C. Three per cent⁵¹Cr release and no complement consumption were observed in VBS-EDTA serum. In contrast, 6%⁵¹Cr release with 37 and 54% decrease in C3 and B hemolytic activities respectively occurred in VBS-MgEGTA serum and 14%⁵¹Cr release with 50% decrease in C2 hemolytic activity occurred in VBS⁺⁺ serum. These results suggest that influenza virus may alter the platelet surface in such a way that both complement pathways might be recruited and the cells be lyzed in autologous serum.The human complement system is activated by a number of viruses and virus-infected cells through antibody-dependent and independent mechanisms. Guinea pig erythrocytes that have been treated with influenza virus are lyzed in human serum through activation of the classical and of the alternative pathways: activation of the alternative pathway is dependent on an acquired resistance of the cell-bound C3 amplification convertase to control mechanisms that are directly related to desialation of the cells by viral neuraminidase [1]. Since,in vivo, clearance of desialated platelets is enhanced in animal models and since human platelets do not express the C3b receptor-associated inhibitory activity of the complement system, we investigated whether human platelets, after contact with influenza virus, acquire the ability to activate complement in autologous serum.     

Clonally restricted insulin autoantibodies in a cohort of 2200 healthy blood donors

Summary Serum samples of 2200 blood donors were screened for anti-insulin IgG by enzyme-linked immunosorbent assay. Specificity of detected antibodies was verified by competition with an excess of insulin and observation that saturated anti-insulin IgG were no longer measurable. The upper limit of measured signal in the population was defined as the mean plus three SD. In the direct assay, 32 sera were positive. Among these, 22 (1%) contained saturable insulin antibodies (true positive) and 10 were non-saturable and considered as non-insulin-specific. The positive blood donors were requested to answer a questionnaire and according to their answers, none had ever received insulin, was a first degree relative of a Type 1 (insulin-dependent) diabetic patient or had experienced a hypoglycaemic episode. None of the 22 true positive sera detected by enzyme-immunosorbent assay bound ¹²⁵-I-insulin to any significant extent. The nine sera yielding the highest signal were further characterized with regard to heavy and light chains as well as species specificity of ligand. Anti-insulin IgG from healthy blood donors contained only one heavy (1 2/9; 3 7/9) and one light ( 8/9; 1/9) chain. Three sera were human insulin specific; two were non-species-specific; the other four bound insulin in the order human = porcine > bovine. These results indicate that low affinity clonally restricted antibodies against insulin are present in unselected blood donors with a prevalence of 1%.     

Efficacy of a support group programme for care-givers of demented patients in the community: a randomized controlled trial

Dementia induces morbidity not only in the patients but also in the families taking care of them. Many studies described the impact of care-giving on physical and psychological health. Support groups were designed to alleviate the burden of care-givers. The objective of this study was to measure the efficacy of a support group programme for care-givers of demented patients in the community. Forty-one primary care-givers were randomly assigned to a study (n=23) or a control group (n=18). Subjects of the study group attended a structured programme of eight 2-h sessions. These weekly sessions consisted of information on the disease, role-playing on management of behavior problems, discussion on emotional impact of care-giving, and learning of stress management techniques. Subjects of the control group were referred to informal monthly meetings of the Alzheimer's Society. Subjects of both groups were evaluated at the entry (T1), after 8 weeks (T2) and after 8 months (T3). The outcome variables were the Burden Interview, the Revised Memory and Behavior Problems Checklist, the Brief Symptoms Inventory, the Alzheimer's Disease Knowledge Test and a questionnaire on health care utilization. Compared with the control group, subjects of the study group yielded only a significant increase in knowledge about the disease (p<0.0001) but no significant difference on the other outcome variables. It is concluded that this type of support group programme seems to have only a minimal impact on morbidity and on the burden of care-givers. These results are similar with two other studies examining the same issue.     

Effets de l’halothane sur la ventilation et les gaz du sang artériel chez le rat avec ou sans paralysie diaphragmatique

Certains patients atteints de paralysie diaphragmatique ou de dysfonctionnement diaphragmatique maintiennent leur ventilation par la mise en jeu d’autres muscles que le diaphragme. L’anesthésie, modifiant le fonctionnement de ces muscles, représente un risque potentiel chez ces patients. Afin d’évaluer ce risque, nous avons étudié les effets de l’halothane sur la ventilation et sur les gaz du sang artériel sur un modèle de paralysie diaphragmatique bilatérale, le rat phrénicectomisé. L’étude a été réalisée sur 43 rats. L’efficacité de la phrénicectomie a été contrôlée par l’observation directe, après laparotomie. La laparotomie n’entraine pas de modification des gaz du sang. Chez 23 rats, une laparotomie a été effectuée et une artère carotide a été cathétérisée. Chez 11 rats témoins, les nerfs phréniques ont été abordés, sans être sectionnés. Chez 12 rats, les phréniques ont été sectionnés. La ventilation a été mesurée par une technique pléthysmographique, chez les rats éveillés, avant et après l’opération, puis chez les mêmes rats anesthésiés avec 1,1%, d’halothane inspiré. Les gaz du sang ont été mesurés après l’opération chez les rats éveillés, puis anesthésiés. Chez les 23 rats opérés on observe, après l’opération, une diminution du poids et de la température centrale, plus importante chez les phrénicectomisés que chez les témoins. Chez les 11 rats témoins, après l’opération, la ventilation augmente, sans modification des gaz du sang. Chez ces rats, l’halothane provoque une diminution de la ventilation minute et de la PaO₂ et une augmentation de la PaCO₂. La phrénicectomie entraine chez les 12 rats, éveillés, une augmentation de la ventilation minute, une hypoxémie et une hypercapnie. Chez ces rats, l’halothane entraine le décès dans trois cas, une diminution de la ventilation minute et une hypercapnie et une hypoxémie importantes chez les neuf autres rats. Les modifications des gaz du sang sont plus importantes que chez les témoins anesthésiés. Chez le rat intact, l’halothane provoque des modifications des gaz du sang comparables à celles observées chez d’autres espèces et chez l’homme. La présente étude confirme les effets de l’halothane sur les muscles respiratoires autres que le diaphragme. Elle met en évidence le risque respiratoire majeur que l’anesthésie peut fair courir aux patients dont la ventilation est maintenue par d’autres muscles que le diaphragme.     

The endothelium-derived relaxing factormediated acetylcholine response of the arterial perfusion pressure after cold storage of the isolated rabbit kidney

The vasodilatation induced by acetylcholine (ACh) in a rabbit isolated perfused kidney was abolished when the tissue was exposed to cold ischemia for 72 h in Euro-Collins (EC) solution. This vasodilatation is due to the release of endothelium-derived relaxing factor (EDRF) from renal vasculature as evidenced by the attenuation following methylene blue pretreatment. When kidneys were preserved in EC solution containing UK 38 485, a thromboxane synthase inhibitor, or nicardipine, a calcium channel blocker, ACh-induced vasodilatation persisted after 72 h of cold ischemia. These results were taken as evidence of tissue protective activity of UK 38 485 and nicardipine and have promising implications for cadaveric kidney transplantation.This paper was presented at the 35th World Congress of the International Society of Surgery in Hong Kong in August 1993     

Retransplantation of the liver after side-to-side caval anastomosis

Caval reconstruction in orthotopic liver transplantation is generally performed by two end-to-end anastomoses, using a portal and caval-axillary bypass to sustain hemodynamic stability. In the piggyback modification, the donor's suprahepatic inferior vena cava (IVC) is anastomosed end-to-side to the recipient's preserved IVC. We have used a recently described variant of the piggyback in 18 patients in whom both IVCs are anastomosed side-to-side. We report two patients who needed three retransplants after this reconstruction and conclude that regrafting can be performed in a quick and safe manner.     

Prevalence of hypovitaminosis a in children of peripheral districts of Campinas São Paulo, Brazil

The prevalence of hypovitaminosis A among children of the peripheral districts of the city of Campinas, S o Paulo, Brazil, was estimated by determining serum retinol levels by High Performance Liquid Chromatography (HPLC) in a sample of 131 children aged between three and ten years, between April 1991 and February 1992. A prevalence of 17.6% and retinol concentrations in the range of 0.35 to 0.70 micromol/L were found (CI=11.1-24.1; 95%), indicating the existence of public health risk Ophthalmological examinations, however, failed to detect any cases of xerophthalmy. Additional characterization of the sample was obtained from 341 children. The per capita income of the average household was surprisingly high for low-income areas. According to FAO-WHO standards, food consumption was adequate only for protein (133.96%). Adequacy levels were low for energy (87.76%) and particularly for vitamin A (66.13%) and iron (42.14%). Height for-age and weight-for-height anthropometric indices revealed that many children were located below -1 standard deviation.     

Surfactant replacement therapy: a multicentric trial comparing two dosage approaches

This is an interim analysis of a multicentric trial that took place in 7 Neonatal Units to compare two initial doses of exigenous pulmonary surfactant (100 mg/kg and 200 mg/kg of phospholipids)by using a porcine surfactant for the treatment of very severe Hyaline Membrane Disease. The initial higher dose produced better oxigenation of arterial blood, reducing the time in high oxygen concentrations,while retreatments were necessary in less than half of the infants receiving 200 mg/kg as initial dose. The protocol of this study allowed the administration of additional doses only when FiO(2) was equal or greater than 0.40 instead of > 0.21, as in a large study recently published, where the same initial doses were given. This resulted in more than 40% reduction in the amount of surfactant administered,with apparently similar clinical outcome.