Razão da contagem de plaquetas ao quadrado/diâmetro do baço-aspartato aminotransferase: avaliação de um novo método não-invasivo para a predição de varizes esofágicas / Platelet count squared/spleen diameter-aspartate aminotransferase ratio: non-invasive method to predict esophageal varices

ABSTRACT BACKGROUND - Variceal bleeding has a high mortality among cirrhotics, and screening with endoscopy is indicated at the diagnosis of cirrhosis. Screening with endoscopy implies discomfort, risks and considerable costs. OBJECTIVE - To evaluate platelet count squared/spleen diameter-aspartate aminotransferase ratio (PS/SA), as a non-invasive predictor of esophageal varices in cirrhotics. METHODS - This cross-sectional study evaluated cirrhotics for PS/SA and presence of esophageal varices. Outpatient records of cirrhotic patients were reviewed for the abovementioned data. Sensitivity, specificity, negative and positive predictive values of PS/SA were calculated. After the univariate analysis, variables with P<0.10 were submitted to a logistic regression. RESULTS - The study included 164 cirrhotics, 59.70% male, with a mean age of 56.7 years. Hepatitis C was the most frequent cause of cirrhosis, being present in 90 patients. Patients were classified as Child-Pugh A in 52.44% and as Child-Pugh B or C in 47.56%. Esophageal varices were present in 72.56% of the patients at endoscopy. PS/SA, with a cutoff of 3x108, had a sensitivity of 95.80% (confidence interval of 95% - 95%CI=0.92-0.99), a specificity of 22.70% (95%CI=0.10-0.35), a positive predictive value of 77.20% (95%CI=0.70-0.84) and a negative predictive value of 66.70% (95%CI=0.42-0.91). In the logistic regression, only platelet count and Child-Pugh score were associated to esophageal varices (P<0.05). CONCLUSION - PS/SA has an excellent sensitivity to predict esophageal varices, allowing almost one fourth of patients without esophageal varices to spare endoscopy. Nevertheless, PS/SA is not independently associated to esophageal varices.

RESUMO CONTEXTO - A hemorragia varicosa tem elevada mortalidade entre cirróticos, e o rastreamento endoscópico de varizes está indicado no momento do diagnóstico da cirrose. O rastreamento endoscópico implica desconforto, riscos e custos consideráveis. OBJETIVO - Avaliar a razão da contagem de plaquetas ao quadrado/diâmetro do baço-aspartato aminotransferase (PQ/BA) como preditor não-invasivo de varizes esofágicas em cirróticos. MÉTODOS - Este estudo transversal avaliou cirróticos quanto ao PQ/BA e à presença de varizes esofágicas. Prontuários ambulatoriais de cirróticos foram revisados quanto a tais dados. Sensibilidade, especificidade e valores preditivos negativo e positivo do PQ/BA foram calculados. Após a análise univariada, variáveis com P<0,10 foram submetidas à regressão logística. RESULTADOS - O estudo incluiu 164 cirróticos, 59,70% masculinos, com média de idade de 56,7 anos. Hepatite C foi a mais frequente causa de cirrose, estando presente em 90 pacientes. Os pacientes foram classificados como Child-Pugh A em 52,44% e em Child-Pugh B ou C em 47,56%. As varizes esofágicas estiveram presentes à endoscopia em 72,56% dos pacientes. PQ/BA, com um ponto de corte de 3x108, teve sensibilidade de 95,80% (intervalo de confiança de 95% - IC95%=0,92-0,99), especificidade de 22,70% (IC95%=0,10-0,35), valor preditivo positivo de 77,20% (IC95%=0,70-0,84) e valor preditivo negativo de 66,70% (IC95%=0,42-0,91). Na regressão logística, apenas a contagem de plaquetas e o escore de Child-Pugh associaram-se às varizes esofágicas (P<0,05). CONCLUSÃO - PQ/BA apresentou excelente sensibilidade para predizer varizes esofágicas, permitido que cerca de um quarto dos pacientes sem varizes esofágicas evitasse a endoscopia. Entretanto, PQ/BA não se associou de maneira independente às varizes esofágicas.

Effect of gold nanoparticles on superoxide dismutase and indoleamine 2, 3-dioxygenase in various rat tissues.

Gold nanoparticles have diverse applications and are being used in food and cosmetic industry, for drug delivery and in the diagnosis and treatment of cancer. However there is a need to study their biochemical mode of action. In this study, in vivo effect of gold nanoparticles on the activities of the two antioxidant enzymes — superoxide dismutase (SOD) and indoleamine 2,3-dioxygenase (IDO) was investigated in various tissues of rats. Rats were injected with 20 μg/kg body wt of 20 nm gold nanoparticles for three consecutive days through intraperitoneal route. The animals were sacrificed by CO2 asphyxiation 24 h after the last dose of gold nanoparticles. Results showed that treatment with gold nanoparticles caused no significant change in SOD activity in most of the tissues, except kidneys. In kidneys, gold nanoparticles caused a significant increase in SOD activity, when compared to the activity in control rats. However, treatment with gold nanoparticles altered the expression pattern of SOD activity in various tissues. For example, in control rats highest SOD activity was demonstrated in heart and least in kidneys and spleen. But, in gold nanoparticles treated rats, maximum SOD activity was observed in liver and the lowest in spleen. Gold nanoparticles caused no significant change in IDO activity in the studied tissues.

use of microencapsulated hepatocytes transplantation reduces mortality and liver alterations in schistosoma mansoni infected hamsters

Hepatocyte transplantation is an attractive therapeutic modality for liver disease as an alternative for orthotropic liver transplantation. The goal of this work was to study the adequacy of intrasplenic hepatocyte transplantation [HCTx] in fresh and microencapsulated forms, in a hamster model of liver fibrosis by Schistosoma mansoni infected hamsters were divided into 6 groups; untreated for 11 weeks [GI] and for 15 weeks [GII] treated with praziquantel [PZQ] 7 weeks PI, and killed 4 weeks [GUI] and 8 weeks [GIV] post-treatment. Treated with PZQ 7 weeks PI, and then treated orally with immunosuppressive drug "cyclosporine [4 weeks post PZQ treatment], 24 hr. before interasplenic injection with fresh hepatocytes [V]. Treated with PZQ 7 weeks PI, and then injected interasplenically [4 weeks post-treatment] with microencapsulated hepatocytes [GVI]. GI and GUI were killed 11 weeks PI for assessment the anti-schistosomal efficacy of PZQ. The other four groups were killed 15 weeks PI for investigation of liver and spleen histology, serum liver enzymes and hepatic oxidative markers before and after HCTx. Freshly isolated hepatocytes with a mean viability 92.9711.2% were used for microencapsulation and transplantation. Histological study showed the presence of transplanted hepatocytes in spleen of recipient. PZQ accelerated healing of hepatic granulomatous lesions as evidenced parasitologically by the increase in the percentage of dead eggs and histologically showing more gfanuloma circumscription with more ova degeneration and less inflammatory cells. The 25-day survival rates in Gil, GIV, GVand GVI were 5/15 [33.3%], 8/15 [53.3%], 10/15 [66.7%] and 9/15 [60%] respecttively. In addition, there were significantly better outcomes in serum biochemical indexes such as ALT, AST, y-GT, ALP, and hepatic SOD and MDA in the fresh and microencapsulated groups than in PZQ-treated group, without great differences between the microencapsulated and the fresh transplanted groups. Liver pathological staining supported these findings

Effects of cocaine on activities of ATPase, LDH and SDH in mouse splenocytes / 法医学杂志

OBJECTIVE@#To examine the effects of cocaine on the activities of ATPase, LDH and SDH in cultured mouse splenocytes in vitro.@*METHODS@#The ATPase, LDH and SDH activities in mouse splenocytes were detected at day 7 after continuous culturing the mouse cells exposed to cocaine hydrochloride in final concentration of 10, 20 and 100 microg/mL in vitro.@*RESULTS@#The activities of ATPase, LDH and SDH in mouse splenocytes exposed to cocaine hydrochloride in final concentration of 10, 20 and 100 microg/mL were significantly decreased after continuous culturing for 7 days.@*CONCLUSION@#The present study demonstrated that cocaine could inhibit the activities of ATPase, LDH and SDH in cultured splenocytes in vitro.

Effects of soluble antigen-induced immune cell activation on steroidogenesis in murine lymphoid organ.

The effect of soluble antigenic (bovine serum albumin, BSA) stimulation to induce steroidogenesis in murine lymphoid organs with concomitant changes in proinflammatory or inflammatory cytokine levels and its implication in the alteration of T-cell response was studied in the mice. Male Swiss albino mice (6-8 weeks old) with average body weight (20 +/- 4 g) were randomly assigned to 3 groups and injected with BSA in presence and absence of Freund's complete or incomplete adjuvant, whereas the control group received only saline. After 3 weeks, animals were sacrificed, and serums as well as lymphoid organs were collected. From the lymphoid tissue homogenate, the activities of steroidogenic enzymes and corticosterone and cytokine levels of the serum were estimated. Steroidogenic enzyme activities in murine lymphoid organs, as well as the pro-inflammatory and inflammatory cytokines levels in serum increased after Freund's complete adjuvant-emulsified BSA administration, as compared to control. The serum corticosterone and serum cytokine profile were also elevated. Results suggested that soluble protein antigen (BSA) administration stimulated steroidogenesis in murine lymphoid tissues and rise in the pro-inflammatory or inflammatory cytokine levels might indicate monocyte recruitment as well as TH1 activation.

Purification and characterization of a type II phosphatidylinositol 4-kinase from rat spleen and comparison with a PtdIns 4-kinase from lymphocytes.

A PtdIns 4-kinase from rat spleen particulate fraction was purified to homogeneity and its molecular properties were compared with a PtdIns 4-kinase from splenic lymphocytes. The enzyme activity was solubilized from spleen particulate fraction with Triton X-100 and chromatographed sequentially on phosphocellulose, DEAE-sephacel, heparin acrylamide and hydroxyapatite columns. The purified enzyme preparation showed a 55 kDa band on SDS-PAGE with silver staining. Renaturation of the enzyme activity from SDS-PAGE showed that it comigrated with the 55 kDa protein. Characterization of the enzyme showed that it was a type II PtdIns 4-kinase. Polyclonal antibodies raised against PtdIns 4-kinase inhibited the enzyme activity in in vitro assays. Analysis of adult rat tissue particulate fractions on immunoblots showed restricted immunoreactivity among PtdIns 4-kinases. However, the immunoreactivity is conserved in lymphoid tissues from mouse to human, suggesting that lymphoid tissue has a distinct PtdIns 4-kinase. Activation of rat splenocytes with Con A showed two fold increase in PtdIns 4-kinase activity. Comparison of PtdIns 4-kinases from spleen and splenic lymphocytes showed identical chromatographic behaviour, molecular mass, immunoreactivity, K(m) values for PtdIns and inhibition by adenosine.

Antioxidant enzyme activities in the lymphoid organs and macrophages of rats trained to perform moderate exercise

Strenous exercise and high levels of athletic competition may suppress immune function, increasing susceptibility to infections. Infections are often associated with a reduction in athletic performance and can have permanent or lethal consequences. Recent research, however; suggests that regular paraticipation in moderate exercise has an immunoenhancing effect but the mechanism involved remains unknown. This study examined the effect of moderate exercise (70 per cent of maximal oxygen consumption - swimming for 1 hour daily at 32 degrees Celsius with 5 per cent body weight extra load attached to the tail) training on antioxidant enzyme activities and lipid peroxidation in the lymphoid organs (mesenteric lymph nodes, thymus and spleen) and macrophages of rats. This modality of physical effort reduced the content of lipide peroxides in the lymphoid organs. The authors assumed that this effect of exercise training resulted in increased activity of antioxidant enzymes: Glutathione peroxidase in the mesenteric lymph nodes (2.1 fold) and spleen (3-fold), catalase in the spleen (5-fold) and Mn-superoxide dismutase (SOD) in the thymus (28 per cent). The exercise training increased the hydrogen peroxide production and phagocytic capacity in macrophages which was accompanied by a higher Mn-SOD activity. Therefore, a moderate exercise may be the able to improve immune function due to changes in the oxidative metabolism of the lymphoid organs and macrophages.

Poly ADP ribosylation as a possible mechanism of microwave--biointeraction.

Electromagnetic fields (EMFs) affect the metabolism of the body including the nervous, endocrine, cardiovascular, hematological as well as the reproductive system. EMFs are environmental pollutants, thus posing a health hazard which can cause steric changes in the molecule located at the cell surface. Microwaves are known to cause chromosomal abberations and act as tumor promoters. The process involves a stream of signals from cell membrane to nucleus and other organelles. The present investigations aim to understand the mechanism of biological effects of microwaves (2.45 GHz). The effect was studied on poly ADP-ribosylation, which is a post translational modification of chromatin protein catalysed by the enzyme poly ADPR polymerase using NAD+ as the substrate. Poly ADP-ribosylation has been shown to be involved in several aspects of chromatin structure and function. Twenty-three days old rats weighing 42-48 gms were exposed at a microwave dose level of 1.0 mW/cm2. After exposure for sixty days the animals were sacrificed and an estimation of poly ADPR polymerase activity was undertaken in different organs of these animals. There was an increase of 20% in its activity in liver, 35% in testis, whereas brain showed a 53% decrease in diencephalon and 20% decrease in the cortex in the exposed animals as compared to their respective controls. There was no change in enzyme activity in spleen and kidney. This was accompanied by concomitant changes in NAD+ levels. The above results may be cited as important events in carcinogenesis and tumor promotion related to microwave exposure and the signal transduction mechanism involved. The goal is to shed light on complex ecogenetic interactions leading to cancer modulation of gene expression by epigenetic mechanism.

5'-Nucleotidase activity in retinol deficiency induced albino rats.

Retinol deficient rat liver, kidney and spleen showed a significant decrease in their enzyme activity (36% to 50%) compared to controls. The lectins could stimulate the enzyme activity in retinol deficient group by 57.6 to 92%, compared to controls (13.3% to 74%). Detergents increased the enzyme activity in retinol deficient tissue microsomes by 4.5%-80% in comparison to controls (10.3% to 119%). The results reveal alterations in membrane structure induced by retinol deficiency.

Effect of aging on the glutaminase activity of neoplastic and immune tissues

1. The effect of age and Walker 256 tumor on maximal phosphate-dependent glutaminase activity of rat immune tissue was determined. Glutaminase is a key enzyme in the metabolism of glutamine, an important fuel for normal and neoplastic cells. 2. Maximal activity of phosphate-dependent glutaminase was measured in immune tissues and tumors of Walker 256 tumor-bearing young (28 days old), mature (3 months old) and aged (15 months old) Wistar rats. The following tissues were examined: thymus, spleen, mesenteric lymph nodes and tumor. 3. Tumor implantation for 14 days reduced glutaminase activity in the thymus and mesenteric lymph nodes. Tumor glutaminase activity was lowest in aged rats and highest in the mature group. 4. Comparison of glutaminase activity in immune and tumor tissues suggested the flux of glutamine between these tissues in the 3 groups. Glutaminase activity was 2.8-fold higher in immune tissues in aged rats (2.58 +/- 0.35 vs 0.93 +/- 0.16 mumol min-1 g tissue wet weight-1, mean +/- SEM, 5 rats), and 1.9- (4.14 +/- 0.47 vs 8.36 +/- 1.29 mumol min-1 g tissue wet weight-1, mean +/- SEM, 5 rats) and 2.5-fold increased (2.41 +/- 0.20 vs 5.92 +/- 0.22 mumol min-1 g tissue wet weight-1, mean +/- SEM, 5 rats) in tumor tissue in the mature and young groups, respectively. These results suggest the deviation of glutamine flux from defense cells to the neoplastic tissue in tumor-bearing young and mature rats and may partially explain the slow cancer growth in elderly patients

Molecular cloning of a membrane bound tyrosine-specific protein kinase from rat spleen.

Tyrosine-specific protein phosphorylation is believed to play an important (though poorly understood) role in various cellular functions in many normal and malignant cells. In order to understand the function of tyrosine-specific protein kinases in normal cells, it is necessary, as an initial step, to identify genes (and proteins) for these enzymes. For this purpose cDNA libraries were constructed in plasmid vector pGEM-3Z and lambda gt11 using mRNA from rat spleen. From these cDNA libraries, cDNA clones coding for a src-related tyrosine-specific protein kinase were isolated. The largest clone (L115) was 1.94 kb in size. Various restriction fragments of this clone were subcloned in plasmid vector for sequencing. The complete nucleotide sequence of the largest clone showed an open reading frame coding for a protein of 503 amino acids. The presence of a glycine at position 2 and an arginine at position 7 indicated that this protein is likely to be acylated at glycine 2 and therefore associated with plasma membrane. This gene showed high homology to human and mouse hck and hence it is perhaps the rat homologue of hck. Moderate level of expression of this gene was observed only in the adult rat spleen and not in other tissues. These results suggest that this kinase gene is expressed in a tissue specific manner.

The effect of malathion dust on certain tissues of male rats fed varying levels of dietary protein.

The effect of malathion, an organophosphorus insecticide on tissue levels of acetylcholinesterase (Ache), phosphomonoesterases and transaminases have been studied in presence of different levels of dietary proteins. Adult male albino rats weighing 150-200 gms were given 5%, 10% and 20% protein diets containing 400 mg malathion (dust) 5% conc./kg feed for 30 days. Its effect was evaluated in liver, kidney, brain, lungs and spleen and results were compared with their respective malathion dust, pair-fed animals (5%, 10% and 20% protein groups without malathion). Animals kept on low protein diets (5% and 10%) when exposed to malathion dust showed significant increase in the activities of GOT and alkaline phosphatase in liver, kidney, brain, lungs and spleen, while a marked inhibition in the activity of Ache was observed under similar treatment. GPT was decreased in kidney and lungs, in the low protein groups (5% and 10%, whereas its activity was increased in liver, brain and spleen of animals receiving 5% protein, when exposed to compared to their respective pair-fed animals. Thus, although the degree of alteration in the enzyme profile is less severe, these changes show that high protein diet has a protective role against pesticide hazards, whereas low protein diet provides less stability to the structural integrity of the tissues.