T Cell-based RAS Activity and Insulin Levels in Obese Subjects with Low Grade Inflammation.

BACKGROUND: Obesity is a major contributor to inflammation and oxidative stress that are key underlying causes for insulin resistance (IR) and diabetes. Accumulated evidence suggest that RAS may serve as a strong link between IR and obesity. We investigated RAS activity in circulating T cells by obese subjects with and without angiotensin (Ang) II stimulation in presence or not of IR and of low-grade inflammation. METHODS: We studied 29 obese and 10 healthy subjects. After T-lymphocytes isolation, mRNAs for angiotensin converting enzyme (ACE) and angiotensin 1-receptor (AT1-R) were quantified by reverse transcription polymerase chain reaction (RT-PCR). High-sensitivity C-reactive protein (hs-CRP), insulin and inflammatory cytokines serum levels, plasma renin activity (PRA) and ACE activity in cell pellet and supernatant, and angiotensin (Ang) II T cell content were also measured. RESULTS: Under baseline conditions, RAS gene expressions, ACE activity and Ang II levels in T cells, but not PRA, of obese subjects with or without IR and with or without hs-CRP ≥3mg/dl were higher than in controls (p < 0.05). The increase in all parameters induced by Ang II was significantly higher in T cells from the obese subjects with hs-CRP ≥3 mg/dl than in controls or in the obese subjects with hs-CRP <3 mg/dl. In the obese subjects with low grade inflammation and IR, the cytokine serum levels and T cells RAS gene expression was inversely correlated with insulin serum concentration. CONCLUSIONS: Low grade inflammation amplifies the T cell RAS response to Ang II stimulation. T cell RAS gene expressions and serum levels of inflammatory cytokines were inversely related with insulin serum concentration. A protective role of insulin towards the development of inflammatory events can be hypothesized.

Pathological Characterization of Lymphoma with Pulmonary Involvement in Cats.

Studies evaluating histological patterns of lung infiltration by lymphoma in cats are lacking. We evaluated retrospectively the histological patterns of 125 necropsy cases of feline lymphoma collected over a 12-year period. Of the 125 cases, 16 showed lung infiltration (12.8%). Nine cats were female, seven cats were male and all were of mixed breed. Gross lesions observed in the lungs consisted of masses (25%) and nodules (18.7%); however, the majority of cases (56.2%) did not show any gross abnormality. Peribronchial-vascular infiltration (93.7%) was the most frequent pattern, followed by pleural (56.2%), interstitial (50%), nodular (37.5%) and alveolar (12.5%) patterns, but 75% of the cases had more than one pulmonary infiltration pattern. There were 14 (87.5%) B-cell and two (12.5%) T-cell lymphomas. Six cats (37.5%) were positive for feline leukaemia virus and three cats (18.7%) were positive for feline immunodeficiency virus. Diffuse large B-cell lymphoma was the most frequent type, accounting for 56% of all cases.

Mortes súbitas em bovinos causadas por Amorimia exotropica no Rio Grande do Sul / Sudden death in cattle due to the consumption of Amorimia exotropica (Malpighiaceae) in Rio Grande do Sul, Brazil

Descrevem-se casos de morte súbita em bovinos associados com a ingestão de Amorimia (Mascagnia) exotropica em seis propriedades rurais localizadas na região metropolitana de Porto Alegre e na serra gaúcha. Os bovinos intoxicados foram encontrados mortos sem história de sinais clínicos prévios, ou apresentaram tremores musculares, quedas bruscas, movimentos de pedalagem, opistótono, respiração ofegante e decúbito lateral, quando induzidos ao movimento poucos minutos antes da morte. Registrou-se maior número de casos entre os meses de maio e agosto. Nove bovinos foram necropsiados e os principais achados macroscópicos observados foram mucosa oral levemente cianótica (3/9), hidropericárdio leve a moderado (3/9), petéquias e equimoses no epicárdio (5/9), coágulo no interior do ventrículo esquerdo (4/9), edema pulmonar (5/9) e mucosas vermelhas no abomaso e no intestino delgado (6/9). Histologicamente havia necrose de coagulação no miocárdio (9/9) caracterizada por retração celular, aumento da eosinofilia do citoplasma com perda das estriações, vacúolos intracitoplasmáticos, núcleos em picnose, vacúolos intranucleares com marginalização da cromatina e ocasionais núcleos em cariorrexia e cariólise. No coração, edema intersticial (3/9) e infiltrado inflamatório intersticial predominantemente mononuclear (7/9) também foram observados. Nos rins de três bovinos havia degeneração hidrópico-vacuolar multifocal das células epiteliais dos túbulos contorcidos distais associada com núcleos picnóticos deslocados para periferia da célula.

Cases of sudden death in cattle were associated with the consumption of Amorimia (Mascagnia) exotropica and occurred in six ranches located in the mountainous region of Rio Grande do Sul and the metropolitan region of Porto Alegre, Brazil. Affected cattle were found dead with no history of previous clinical signs, or showed muscular tremors, falls, paddling, opistotonus, panting, and lateral recumbence after being induced to move, few minutes before death. Most cases were recorded between May and August. Nine cattle were necropsied and main gross findings were oral mucosae slightly cyanotic (3/9), mild to intermediate hydropericardium (3/9), epicardial petechiae and ecchymoses (5/9), clot within the left ventricle (4/9), lung edema (5/9), apart of abomasal and small intestinal reddened mucosa (6/9). Histologically, there was myocardial coagulation necrosis (9/9), which was characterized by cellular retraction, enhanced cytoplasmic eosinophilia, lack of cytoplasmic striations, intracytoplasmic vacuoles, nuclear piknosis, intranuclear vacuoles, chromatin marginalization and occasional nuclear karyorrhexis and karyolysis. There also were interstitial edema (3/9) and interstitial inflammatory infiltrate (mainly mononuclear) (7/9) in the heart, apart of multifocal vacuolar-hydropic degeneration in the epithelial cells of the distal convoluted tubules associated with pyknotic and eccentric nuclei in the kidneys of three cattle.

Expression of measles virus antigens in Streptococcus gordonii.

The measles virus proteins haemagglutinin (HA) and fusion protein (F), which together mediate attachment and penetration of the virus in the host cell and can elicit production of neutralising antibodies in the course of natural infection were expressed in the vaccine vector Streptococcus gordonii, a Gram-positive bacterium normally present in the human oral cavity. HA and F were expressed as fusion proteins attached to the bacterial surface, and were both found to be immunogenic when the recombinant S. gordonii were inoculated subcutaneously in mice.

[Neuroleptic malignant syndrome: a neurologic pathology of great interest for the internist]. / Sindrome maligna da neurolettici: una patologia neurologica di grande interesse per l'internista.

Two cases of neuroleptic malignant syndrome are reported. Although affecting chiefly psychiatric patients, this rare, polymorphic disease should be well-known to internists, general practitioners, and emergency staff because of its high risk of fatality. Our patients, who presented without some of the main symptoms, fell in the category of incomplete variants of the syndrome, the most difficult to recognize. In both patients, computerized tomography carried out at the outset of symptoms, evidenced signs of cerebral edema, a datum never before reported. The symptomatic polymorphism, pathogenesis, differential diagnosis, and therapy of this disease are illustrated and discussed.

Detection of human herpesviruses 6 and 7 in heart transplant recipients by a multiplex polymerase chain reaction method.

In order to evaluate the possible reactivation of human herpesviruses 6 (HHV-6) and 7 (HHV-7) after heart transplantation, buffy-coat and plasma specimens from 21 transplant patients and 56 healthy blood donors were examined for HHV-6 and HHV-7 DNA by polymerase chain reaction. Human herpesvirus 6 and HHV-7 infection or reactivation has been suggested to play a role in cytomegalovirus disease progression in renal transplant recipients. In the present study, however, no significant difference in the prevalence of HHV-6 and HHV-7 was found between the immunosuppressed and the healthy population; moreover, no viral reactivation was found in the heart transplant recipients.

Response to measles revaccination among Bolivian school-aged children.

The response to measles revaccination was evaluated in 1994 among 202 Bolivian school-aged children whose antibody levels were below 200 miu (milli-international units) by haemagglutination inhibition (HI) in a large-scale serosurvey conducted in Santa Cruz one year earlier. Of the 202 revaccinated children, 164 (82%) had seroconverted between the 1993 serosurvey and the pre-revaccination blood sample. A measles outbreak occurred in Santa Cruz 6 months before the revaccination. Among the seroconvertors, only 6% gave a history of measles, and 15% a history of contact with a case of measles. All 20 children with undetectable HI antibody pre-revaccination, and all 6 children with levels below 100 miu, seroconverted after revaccination. The geometric mean titres by HI at 4 weeks after revaccination were 2018 miu (95% confidence limits [95% CL] 1143, 3564) and 398 miu (95% CL 254, 625) in the 2 groups, respectively. Six of 9 children with pre-revaccination antibody titres of 100-199 miu also seroconverted. No child demonstrated a measles-specific immunoglobulin M response. Among the 29 children who seroconverted and were followed up at one year after revaccination, 15(52%) showed a fourfold or greater decline in antibody levels, which in 8 fell to levels below 200 miu. This study confirmed the observation that revaccination is successful in producing an antibody response in children with low or undetectable pre-revaccination titres, but it also confirmed that vaccine-induced immunity wanes rapidly.

Prevalence of measles antibody among children under 15 years of age in Santa Cruz, Bolivia: implications for vaccination strategies.

We conducted a community-based survey in Santa Cruz city, Bolivia, to determine the age-specific prevalence of measles antibodies, determine factors associated with absence of detectable measles antibodies, and to compare results of salivary and serum measles immunoglobulin G (IgG) antibody assays. Serum samples from 1654 children were assayed for measles IgG antibody using the haemagglutination inhibition (HI) assay, and salivary samples were also obtained from 187 children and tested for measles IgG antibody using an antibody capture radioimmunoassay. Reported measles vaccine coverage in children aged 12-35 months was 77% (95% confidence interval [CI], 72-81%). Eighty-seven percent (95% CI 85-89%) had detectable HI antibody, but a high proportion had antibody levels below 200 miu (30-40% of 2-14 years old children). Measles seronegativity was associated with not being vaccinated against measles, a negative history of measles disease, living in the inner city, being a lifetime resident of Santa Cruz, and young age. Of 212 children without detectable measles antibody, 58% had a positive history of vaccination or measles disease, so that historical information was not sufficiently reliable to identify susceptibles. The salivary measles antibody assay was not sufficiently sensitive to be used for population screening; only 54% of 171 salivary samples from children who had detectable serum HI antibody were positive. A mass measles vaccination campaign of all children under 15 years of age is planned in Bolivia in 1994. Although only 7% of school-age children in Santa Cruz were seronegative, the effectiveness of a mass campaign in this age group depends in part on the response to revaccination of children with low, but detectable, antibody levels.

Prevalence of measles antibodies before and after vaccination in previously unvaccinated children of the Cordillera Province (Santa Cruz Department, Bolivia).

A study was carried out in the Cordillera Health District (Santa Cruz Department, Bolivia) from October 1988 to April 1989 to determine the seroprevalence of measles antibodies and the seroconversion rates among a group of previously unvaccinated children (9-36 months of age) from the urban and rural area of the province, before and after immunization with a standard dose of Schwarz measles vaccine. Among 265 previously unvaccinated children, 77 (29%) had measles IgG antibodies prior to immunization; 141 out of 147 (96%) seronegative children at the time of vaccination seroconverted. No difference in seroprevalence and in seroconversion rates was found between the urban and rural groups.

Nuclear magnetic resonance investigation of virus-lymphomonocyte interactions.

The water protein spin-lattice relaxation time (T1) was measured in suspensions of human peripheral blood mononuclear cells (PBMC), uninfected or infected with type 1 herpes simplex virus, human cytomegalovirus and rubella virus. In the infected samples, T1 enhancements, which linearly depend on virus concentration, were observed. This T1 increase can be related to the early changes induced by the virus adsorption of the cells, not always confirmed by virus-induced cytopathic effect (CPE) in cocultures of infected PBMC and other sensitive cells. Compared with other conventional virological techniques, this NMR method seems to be rapid and sensitive. The NMR response was reproducible and specific, since neutralization of the viral infection by homologous antisera consistently matched the neutralization of the virus-induced NMR effects. These observations suggest that fast and sensitive 1H-NMR relaxation techniques can be implemented in virological diagnosis directly on pathological materials.

Anthracycline cardiotoxicity: in vivo and in vitro effects on biochemical parameters and heart ultrastructure of the rat.

The premise of this study is that mitochondrial lesions caused by anthracyclines lead directly to cardiotoxicity. We compared several biochemical parameters, including endogenous cellular respiration, adenosine and guanosine triphosphate levels, and 14C-amino acid incorporation, of rat hearts treated with doxorubicin and some of its derivatives, recent products of pharmacological research aimed at selecting less toxic antiblastic agents. In rats treated in vivo, we further examined the ultrastructural changes induced by anthracycline antibiotics in order to elucidate which biochemical parameters were consistent with the morphological lesions. Our data indicate that mitochondria are the target of the anthracycline effects and that oxygen uptake and nucleotide levels may be regarded as markers of the toxicity when evaluating new drugs before their clinical use. The lack of cytoplasmatic or endoplasmatic reticulum alterations may account for the failure of anthracyclines to affect amino acid incorporation. In any event, the rate of protein synthesis cannot serve as a marker of cardiac toxicity. In this context, epidoxorubicin and iododoxorubicin are two derivatives characterized by less cardiotoxic potential than doxorubicin and thus appear to be promising antiblastic agents.

Sarcoidosis and infections by human lymphotropic viruses.

In sera of 38 patients with sarcoidosis, assayed for antiviral antibody to Rubella, Measles, Cytomegalovirus (CMV), adenovirus, Epstein-Barr virus (EBV), human immunodeficiency virus (HIV), different incidence of seropositivity have been found, according to the virus assayed. Interestingly all patients were seropositive to rubella virus with high titres compared to data obtained from normal age matched population. These data indicate that in Sarcoidosis an altered antibody response of post-primary type can be found.

NMR investigation of cell cultures: early detection of infection by Herpes simplex virus type 2 and transformation.

The water proton Nuclear Magnetic Resonance (NMR) spin-lattice relaxation rates were measured in suspension of uninfected and Herpes simplex virus type 2-infected WI-38 diploid cells and shown to allow reliable detection of (i) the early signs of productive infection, and (ii) the tendency to undergo either spontaneous or virus-induced transformation. In addition, NMR investigation of several transformed cell lines currently and previously established from WI-38 cells clearly indicated that the NMR relaxation parameter is a suitable tool for assessment of the level of malignancy, as revealed by in vivo oncogenicity tests in immunosuppressed mice.

Live-cell immunofluorescence for detection of human immunodeficiency virus type 1 (HIV-1)-related antigens in peripheral blood mononuclear cells from HIV-1 seropositive individuals.

Live-cell indirect immunofluorescence was used to detect HIV-associated antigens in fresh uncultured peripheral blood mononuclear cells from 29 HIV-seropositive subjects. Conventional fixed-cell immunofluorescence and live-cell immunofluorescence were run in parallel on each sample using monoclonal antibodies directed against HIV gag and env gene products. Sera from the same patients were also tested for HIV antigen by a sandwich enzyme immunoassay. Results strongly indicate that nondenaturing live-cell immunofluorescence is much more sensitive for detection of HIV-associated antigens and may be useful in the diagnosis of HIV infection as well as in the study of pathogenesis.

Doxorubicin and epirubicin cardiotoxicity: experimental and clinical aspects.

Epirubicin (EpiDx) belongs to the class of anthracycline antibiotics. It is an analog of doxorubicin (Dx) modified in the sugar moiety and in which the stereochemistry at the hydroxyl group bearing C-4' has been inverted. The purpose of this study was to evaluate in an experimental model and in a clinical trial the cardiotoxic effects of EpiDx vs Dx. Cellular oxygen uptake was measured in vitro for equal concentration of Dx and EpiDx with a Warburg manometric apparatus and ATP intracellular concentration by high-pressure liquid chromatography. Dx inhibits cellular endogenous respiration of rat heart slices by 34% vs control, while EpiDx reduces oxygen uptake by 18%. ATP intracellular concentration was significantly reduced by both anthracycline derivates. The same results were obtained after Dx administration in vivo which enabled us to correlate the biochemical parameter (QO2) with the histological cardiac damages shown by light microscopy. For the clinical trial, we studied a total of 22 patients undergoing chemotherapy for solid tumors in an advanced stage. Nine of these were treated with Dx for a total of 66 therapeutic cycles and reached a maximal cumulative dose of 540 mg/m2. Thirteen were treated with EpiDx for a total of 121 cycles (maximal dose reached 720 mg/m2). The dosage of both agents were equal, 60 mg/m2 every 3 weeks. Acute cardiotoxicity was evaluated, measuring creatine-kinase isoenzyme MB(CK-MB) serum level before and 15 h after anthracyclines administration.(ABSTRACT TRUNCATED AT 250 WORDS)

[Evaluation of the specific immune response after anti-rubella vaccination]. / Valutazione della risposta immunitaria specifica dopo vaccinazione anti-rosolia.

A selective rubella immunization program was carried out in the Siena (Italy) area since 1984. 912 schoolgirls aged 10 to 18 were screened for rubella antibodies. Seropositivity rates, increasing with age, appeared quite heterogenous among subjects living in different villages. Seronegative girls were partly voluntarily immunized with RA 27/3 live attenuated rubella vaccine. Serological and clinical controls performed on 238 subjects one month after immunization showed a 99.6% vaccine-induced seroconversion rate and mild post-vaccinal reactions in 26% of vaccinees. Serological control performed, on 172 subjects, at 8-12 days after vaccination revealed that in most cases vaccine induced haemagglutination-inhibiting antibodies are not detectable at the time. The results are discussed in relation to a forthcoming introduction of compulsory rubella vaccination in our Country.