Antibodies to M-type phospholipase A2 receptor (PLA2R) in membranous lupus nephritis.

BACKGROUND: Antibodies to M-type phospholipase A2 receptor (a-PLA2R) have been identified in most patients with idiopathic membranous nephropathy, but the prevalence in membranous lupus nephritis (MLN) is still unclear. The objective of this study was to assess the prevalence of a-PLA2R antibodies in a large cohort of patients with lupus nephritis. METHODS: a-PLA2R antibodies were measured by ELISA in serum from patients with systemic lupus erythematosus ( n = 190), of whom 37 had a biopsy-proven MLN. Positive samples were confirmed by commercial ELISA kit, Western blot and immunohistochemistry in renal tissue. RESULTS: A total of 10 from 190 patients (5.3%) with systemic lupus erythematosus had circulating a-PLA2R measured by in-house ELISA assay. The antibodies were detected in 7 patients with MLN (18.9%) and 3 patients with non-renal lupus disease (3.2%). PLA2R staining was detected in the kidney biopsy of 5 of the 7 (71.4%) patients with MLN. a-PLA2R levels were associated with active disease but not proteinuria levels. Presence of a-PLA2R antibodies at baseline was associated with worse remission rates and longer time to remission compared to those patients serologically negative. CONCLUSIONS: a-PLA2R antibodies can be detected with low prevalence in MLN patients, but their detection is associated with a worse renal prognosis.

The Mexican consensus on the treatment of hepatitis C. / Consenso Mexicano para el Tratamiento de la Hepatitis C.

The aim of the Mexican Consensus on the Treatment of HepatitisC was to develop clinical practice guidelines applicable to Mexico. The expert opinion of specialists in the following areas was taken into account: gastroenterology, infectious diseases, and hepatology. A search of the medical literature was carried out on the MEDLINE, EMBASE, and CENTRAL databases through keywords related to hepatitisC treatment. The quality of evidence was subsequently evaluated using the GRADE system and the consensus statements were formulated. The statements were then voted upon, using the modified Delphi system, and reviewed and corrected by a panel of 34 voting participants. Finally, the level of agreement was classified for each statement. The present guidelines provide recommendations with an emphasis on the new direct-acting antivirals, to facilitate their use in clinical practice. Each case must be individualized according to the comorbidities involved and patient management must always be multidisciplinary.

[The impact of antimicrobial resistance and Streptococcus pneumoniae serotype distribution on the mortality of children under 5 years of age with invasive disease]. / Impacto de la resistencia a antimicrobianos y de serotipos de Streptococcus pneumoniae en la mortalidad de niños menores de 5 años con enfermedad invasora.

Severe pneumonia and meningitis caused by Streptococcus pneumoniae have been persistently associated with high mortality rates, despite advances in antimicrobial therapy and the development of vaccines. Resistance to penicillin and other antimicrobial agents is increasing and spreading worldwide. Even though risk factors for development of antimicrobial resistance have been identified, their influence on mortality has not been clarified. With regard to virulence, differences among serotypes have been determined, but their impact on mortality is unknown. The aim of this study was to determine the risk factors associated with mortality in children with invasive pneumococcal disease. Clinical records for 245 children under 5 years of age with invasive disease due to S.pneumoniae were reviewed. Children were diagnosed between 1994 and 1996 in Colombia, during the study of S.pneumoniae capsular types conducted by the Pan American Health Organization's Regional System for Vaccines. Of the 245 patients whose charts were examined, 29 (11%) died. No significant differences in age, gender, underlying disease, nor antimicrobial treatment concordance were found. Variables associated with mortality in the univariate analysis were a diagnosis of meningitis; antimicrobial resistance to penicillin, trimethoprim-sulfamethoxazole (TMS), or erythromycin; multiresistance, and serotypes 6, 23F, 7F, 8, and 35B. In the logistic regression, serotypes 7F (OR = 7.13; P = 0.04) and 8 (OR = 13.8; P = 0.07), polipnea (OR = 2.74; P = 0.03), meningitis (OR = 5.02; P = 0.0001) and TMS resistance (OR = 2.62; P = 0.02) continued to be associated with mortality. In patients with pneumonia, serotype was the factor most consistently associated with mortality; in meningitis patients, it was antimicrobial resistance. Differences in mortality according to serotype must be taken into account in developing a vaccine if a substantial impact on pneumococcal disease morbidity and mortality is to be achieved.

Repeated doses of the perfluorocarbon FC-100 improve lung function of preterm lambs.

Intratracheal administration of a single dose of the perfluorocarbon FC-100 improves lung function in surfactant-deficient animals. In this study we compared the response to repeated doses of FC-100 (3 mL/kg 3% solution, n = 5) with that observed after administration of Exosurf (5 mL/kg, n = 5) to mechanically ventilated preterm lambs of 125 d of gestation. The initial dose of FC-100 rapidly increased arterial PO2, decreased arterial PCO2, and improved arterial pH. Also dynamic lung compliance markedly improved with this agent. Administration of an additional dose of FC-100 resulted in relatively similar changes, albeit of lesser magnitude than those observed with the initial dose. In contrast, Exosurf did not improve these variables even after three doses. All lambs treated with FC-100 survived the 6-h study period, whereas one of the five Exosurf-treated lambs survived (p < 0.05). Mean arterial blood pressure and heart rate decreased in those lambs that received FC-100, but not in surviving lambs that received Exosurf. Our data demonstrate that repeated intratracheal administration of the perfluorocarbon FC-100 improves lung function and survival of surfactant-deficient lambs better than the synthetic surfactant Exosurf. We speculate that tensio-active agents with properties different from surfactant, such as FC-100, might improve lung function in preterm neonates with diseases due to surfactant deficiency.

Uso do probiótico "Paciflor" em dieta para coelhos em crescimento / The use of probiotic \"Paciflor\" in diet for growing rabbits

Com o objetivo de avaliar-se a eficácia nutricional do probiótico Paciflor R, cujo princípio ativo está constituído por esporos de Bacillus IP 5832, alojaram-se, individualmente, 60 coelhos de ambos os sexos, dos 30 dias de idade até alcançarem 2 quilos de peso vivo, alimentados "ad libitum" com duas dietas peletizadas e equitativamente equilibradas com 16 por centro de proteína bruta, 14 por cento de fibra bruta e 2500 kcal de energia digestível por quilo de dieta. Em uma das dietas foi incluído o probiótico na concentraçäo de 10 elevado à sexta potência esporos por grama. Ao final do período de crescimento, sacrificaram-se 44 coelhos dos dois grupos experimentais para avaliaçäo do rendimento de carcaça e dos pesos totais das vísceras e do ceco repletos de digestiva. Os resultados mostraram que a inclusäo do paciflor R näo alterou o período de crescimento (média = 48,4 dias) e o ganho de peso médio diário (média = 30,2 gramas), mas reduziu o consumo médio diário (102,1 vs 119,1 gramas) e a conversäo alimentar (3,5:1 vs 3,9:1). A mortalidade ocorrida no experimento näo foi associada ao tratamento com o probiótico. O rendimento de carcaça (média = 62,2 por cento) näo foi modificado, assim como as proporçöes dos pesos totais das vísceras e cecos repletos de digesta em relaçäo ao peso vivo (média = 16,4 por cento e 4,9 por cento respectivamente). Conclui-se que a inclusäo do probiótico Paciflor R em dietas compostas para coelhos em crescimento pode representar uma economia importante nos gastos com alimentaçäo sem, no entanto, induzir alteraçöes no ganho de peso e rendimento de carcaça

[The utilization of health services and the motives for consultation as indicators of family dysfunction]. / La utilización de servicios de salud y los motivos de consulta como indicadores de disfunción familiar.

OBJECTIVE: To analyse the relationship between the reasons for seeking care on demand and use of the health services on the one hand and disturbances in the family dynamic, on the other, with an attempt to determine indicators of family dysfunction. DESIGN: Crossover descriptive study where the sample was obtained by systematic sampling. The family APGAR test was applied to the whole sample, along with an analysis of the use of services and the reasons for attendance. SETTING: Almanjayar Health Centre during the first 6 months of 1992. PATIENTS: 356 patients over 18. MEASUREMENTS AND MAIN RESULTS: We confirmed that the likelihood of belonging to a dysfunctional family was 77%, with a risk of 6.5 for hyper and normal users. Regarding reasons for seeking care, those who attended because of ill-defined signs and symptoms are 6.21 times more likely to present a family dysfunction than the rest of those interviewed. Patients with ill-defined symptoms or psychiatric disorders who are hyper-users are 7.20 times more likely to present some family dysfunction. CONCLUSIONS: It will be useful to carry out a study of family function among hyper-users, those who attend because of ill-defined signs and symptoms and people with mental health problems.

[Development of local health systems in Uruguay]. / Desarrollo de los silos en el Uruguay.

This article summarizes Uruguay's experience in local health systems development and the elements that have contributed to awareness and dissemination of this concept. In 1988 the Ministry of Public Health assumed responsibility as one of the entities charged with strengthening the country's local health systems. A technical group was created to act at the central level. It has proposed and promoted changes directed toward the deconcentration of resource utilization in accordance with a set of general guidelines and, at the local level, has acted as a catalyst in the understanding of health services delivery management as a systemic concept.

CD9 is an unreliable antibody for bone marrow purging.

Various procedures are useful in purging bone marrow of pre-B lineage contamination prior to autologous bone marrow transplantation. Specific monoclonal antibodies are used in combination with complement, magnetic microspheres, chemotoxins, phototherapy or lectins. In this setting, the usual monoclonal antibody cocktail consists of CD9 (DU-ALL-1), CD10 (WCMH 15.14) and CD19 (HD37). Antigens reacting with all three monoclonal antibodies are present on early and mature pre-B lymphocytes. However, CD9 positive antigens have also been shown to be present on megakaryocytes, platelets and mature granulocytes. In our studies, we have found that CD10 and CD19 expression predictably vary according to disease status, with low positivity in remission and higher positivity in relapse. CD9 expression, however, varies independently of disease status and is frequently inconsistent within individual patients. As such, use of CD9 for marrow purging may result in the removal of a broad range of cell populations not related to the underlying disorder.

Immunomagnetic microsphere mediated purging of cALLa positive leukemic cells from bone marrow for autologous reinfusion.

Bone marrow from sixteen patients with cALLa positive ALL have been treated with a monoclonal antibody (MoAb) cocktail which includes DuALL-1 (CD9), WCMH15.14 (CD10) and HD-37 (CD19). Following antibody treatment, marrows were incubated (30 minutes) with anti-murine-IgG1 (Fc) coated magnetic microbeads and passed through a graded magnetic field chamber. Two problems have had to be addressed: (1) More marrow cells must be processed than in marrows from patients with neuroblastoma, for which the separation chamber was originally developed, requiring the use of more beads, causing occlusion of the chamber's collection surface. This has been corrected by using a large surface area pre-magnet for the elimination of excess microbeads prior to processing in the main chamber; (2) Up-modulation (up to 40% positive cells) of the CD9 associated antigen has been detected. To avoid difficulty, marrows are being routinely screened prior to harvest for purging, and purging is delayed for patients with elevated CD9 levels. Eight patients have been reinfused, with no morbidity or mortality associated with the purging or other ex vivo handling of the marrow.

Lipid coating of paramagnetic microspheres reduces non-specific binding to Kelly neuroblastoma cells.

In an effort to reduce non-specific binding interactions (binding in the absence of antibody), sheep anti-mouse IgG1 (Fc) antibody linked to magnetic microspheres (also referred to as microbeads or beads) were treated with phosphatidylcholine (PC), phosphatidylglycerol (PG), or a mixture of PC and PG. The lipid-treated microspheres were mixed with Kelly neuroblastoma cells, which had been pre-labelled with Hoechst fluorochrome. After 30 minutes of incubation, the microsphere adherent cells were separated from the non-adherent cells and counted. PC or mixed lipid treatment of beads reduced nonspecific binding to 8.3%, compared to 25.8% in the untreated samples. PG, on the other hand, increased non-specific binding. Lipid treatment of beads did not adversely affect specific antibody mediated binding. When a non-specific antibody was added to the incubation mixture, non-specific binding of untreated control beads to cells was increased, but binding of PC-treated beads was unaffected.

Non-specific cell binding characteristics of para-magnetic polystyrene microspheres used for antibody-mediated cell selection.

The binding of cells to paramagnetic polystyrene microbeads in the absence of coupling antibodies was measured. Cells from normal bone marrow, from an acute lymphoblastic leukemia (ALL) cell line or from a neuroblastoma cell line, were labeled with the fluorochrome Hoechst 33342 and incubated with microbeads by rotation at 4 degrees C. Following this incubation, the microbeads with all attached cells were collected using an externally applied magnetic field and visualized by microscopic examination under ultraviolet illumination. The incubation variables included the protein content of the medium, and the period of rotation. Normal bone marrow was found to adhere sparingly to the microbeads; less than 1.0% of the total nucleated cell population was recovered with the beads, whereas greater than 5% of the ALL cells and greater than 30% of the neuroblastoma cells were found to bind non-specifically to the microspheres. Neither changes in the protein concentration of the medium or in the incubation period significantly altered the non-specific binding of the cell types examined. It is thus apparent that the use of these microspheres for positive selection of cells, such as the collection of hematopoietic stem cells for transplantation, would be compromised by a sizeable non-specific interaction. Modification of the surface of the microspheres to substantially reduce this interaction will be necessary before efforts at positive selection using the magnetic microspheres can be fruitful.

Clinical pharmacology of the biological response modifier maleic anhydride divinyl ether copolymer (MVE-2).

The anionic pyran copolymers represent a novel class of high molecular weight biological response modifiers with antitumor activity. Clinical pharmacology studies were performed on MVE-2, a polymer with an average molecular weight of 15.5 Kd. MVE-2 was analyzed in plasma and urine by HPLC. In addition, pharmacology studies were also performed using [14C] labeled MVE-2. The clearance of unlabeled MVE-2 from plasma was monophasic and the t1/2 for MVE-2 was extremely short (between 10 and 26 min). The apparent volume of distribution (Vd) varied from 12-18 l. Both the t1/2 and the Vd did not appear to be dose-dependent. The plasma clearance for [14C] labeled MVE-2 was studied in seven patients. The clearance of [14C] labeled MVE-2 fit a biphasic mathematical model. The alpha phase half-life was between 11 and 18 min while the beta phase half-life was between 70 and 85 min. Urinary excretion for either unlabeled drug or the [14C] label was between 30 and 45% of the administered dose. These studies show that, in man, the polyanionic macromolecule MVE-2 is cleared rapidly from plasma and excreted extensively in urine.

Characterization of tumor-associated lymphocytes in a series of mouse mammary tumor lines with differing biological properties.

Tumor-associated lymphocytes were isolated by isokinetic gradient separation from five related mouse mammary tumor lines with different immunological and growth characteristics. Although considerable variation in recovery rates was seen from experiment to experiment, the five tumor types were found to have reproducible and characteristic patterns of T lymphocyte subpopulations, as detected by cytotoxicity assay using monoclonal antisera to Thy-1, Lyt-1, and Lyt-2 antigens. Tumors of line 168, which are weakly immunogenic at best, had the lowest numbers of recovered ALS+, Thy1+ lymphocytes (12% and 9%, respectively), in contrast to immunogenic lines (mean 38% and 26%, respectively). Line 68H tumors, which grow after prolonged latency periods and also produce tumor cell variants in vivo, were unique in that the numbers of recovered Lyt 1+ lymphocytes exceeded the number of Lyt 2+ lymphocytes, whereas these two T cell subpopulations were either equal or Lyt 2+ cells predominated in the other faster growing, non-variant-producing tumors. No differences in T lymphocyte distribution were associated with the presence or absence of metastatic behavior. These results indicate that distinctive lymphocyte infiltrates may be characteristic of tumors with distinct biological differences.