Desordens hematológicas em onças-pardas (Puma concolor; Linnaeus, 1771) infectadas por Citauxzoon felis / [Hematological disorders in Brown jaguars (Puma concolor; Linnaeus 1771) infected by Citauxzoon felis]

Este estudo objetivou descrever o aspecto hematológico de seis onças-pardas (Puma concolor) infectadas pelo Cytauxzoon felis. Os seis casos de infecção foram identificados durante o manejo sanitário de 11 animais de um centro de reabilitação de animais silvestres. Estruturas compatíveis com piroplasmídeos foram observadas durante a avaliação do esfregaço sanguíneo e confirmadas como Cytauxzoon felis pela técnica de PCR. A análise estatística demonstrou diferença significativa (P<0,05) no número absoluto dos linfócitos entre os grupos dos animais infectados e não infectados. Assim, expressivas alterações hematológicas e bioquímicas entre os grupos investigados alertam para a dificuldade de identificação de onças-pardas infectadas por C. felis, apoiada apenas em exames de rotina, bem como para o risco, sobretudo, da reintrodução desses animais na natureza.(AU)

This Cytauxzoon felis by the PCR technique. Statistical analysis showed a significant difference is study aimed to describe the hematological appearance of six puma (puma concolor) infected with cytauxzoon felis. The six cases of infection were identified during the sanitary management of 11 animals from a wild animal rehabilitation center. Piroplasmid compatible structures were observed during the blood smear evaluation and confirmed as (P<0.05) in the absolute number of lymphocytes between the groups of infected and uninfected animals. Thus expressive hematological and biochemical alterations between the groups investigated alert to the difficulty of identifying infected brown jaguars by C. felis, supported only by routine examinations, and the risk especially when aiming at the reintroduction of these animals in the wild.(AU)

Cytotoxic, clastogenic and genotoxic effects of cis-tetraammine(oxalato)ruthenium(III) dithionate on human peripheral blood lymphocytes

There is a concern about stablishing the clinical risk of drugs used for cancer treatment. In this study, the cytotoxic, clastogenic and genotoxic properties of cis-tetraammine(oxalato)ruthenium(III) dithionite - cis-[Ru(C2O4)(NH3)4]2(S2O6), were evaluated in vitro in human lymphocytes. The mitotic index (MI), chromosomal aberrations (CA) and DNA damage by comet assay were also analyzed. The MTT test revealed that the ruthenium compound showed a slight cytotoxic effect at the highest concentration tested. The IC50 value for the compound after 24 hours of exposure was 185.4 µM. The MI values of human peripheral blood lymphocytes treated with 0.015, 0.15, 1.5 and 150 µM of cis-[Ru(C2O4)(NH3)4]2(S2O6) were 6.1, 3.9, 3.2 and 0.2%, respectively. The lowest concentration, 0.015 µM, did not show any cytotoxic activity. The CA values for the 0.015, 0.15 and 1.5 µM concentrations presented low frequency (1.5, 1.6 and 2.3%, respectively), and did not express clastogenic activity when compared to the negative control, although it was observed clastogenic activity in the highest concentration tested (150 µM). The results obtained by the comet assay suggest that this compound does not present genotoxic activity at lower concentrations. The results show that cis-[Ru(C2O4)(NH3)4]2(S2O6) has no cytotoxic, clastogenic or genotoxic in vitro effects at concentrations less than or equal to 0.015 µM. This information proves as promising in the treatment of cancer and is crucial for future trials.

Platelet/lymphocyte ratio and mean platelet volume in patients with granulomatosis with polyangiitis

Abstract Background: Granulomatosis with polyangiitis (GPA) is a granulomatous necrotizing vasculitis with high morbidity and mortality. Anti-neutrophil cytoplasmic antibody is a valuable diagnostic marker, however its titer lacks predictive value for the severity of organ involvement. Platelet to lymphocyte ratio (PLR) and mean platelet volume (MPV) has been regarded as a potential marker in assessing systemic inflammation. We aimed to explore the value of PLR and MPV in the assessment of disease activity and manifestations of disease in GPA. Methods: 56 newly diagnosed GPA patients and 53 age-sex matched healthy controls were included in this retrospective and cross-sectional study with comparative group. Complete blood count was performed with Backman Coulter automatic analyzer, erythrocyte sedimentation rate (ESR) with Westergen method and C-reactive protein (CRP) levels with nephelometry. The PLR was calculated as the ratio of platelet and lymphocyte counts. Result: Compared to control group, ESR, CRP and PLR were significantly higher and MPV significantly lower in GPA patients. In patients group, PLR was positively correlated with ESR and CRP (r = 0.39, p = 0.005 and r = 0.51, p < 0.001, respectively). MPV was negatively correlated with ESR and CRP (r = - 0.31, p = 0.028 and r = - 0.34 p = 0.014, respectively). Patients with renal involvement had significantly higher PLR than patients without renal involvement (median:265.98, IQR:208.79 vs median:180.34 IQR:129.37, p = 0.02). PLR was negatively correlated with glomerular filtration rate (r = - 0.27, p = 0.009). A cut-off level of 204 for PLR had 65.6% sensitivity and 62.5 specificity to predict renal involvement. Conclusion: PLR exhibit favorable diagnostic performance in predicting renal involvement in patients with GPA.(AU)

Analysis of neutrophil-lymphocyte ratio as a prognostic element in the response to neoadjuvant therapy in rectal cancer / Análise da relação neutrófilo-linfócito como elemento prognóstico na resposta à terapia neo-adjuvante no câncer do reto

INTRODUCTION: The previous radio-chemotherapy approach is highly relevant in the management of rectal cancer, collaborating on organ functional preservation, being performed prior to surgery. The inflammatory response plays an important role in this treatment. OBJECTIVE: It consists in correlating the number of peripheral lymphocytes and the neutrophil/lymphocyte ratio in the peripheral blood with tumor response to neoadjuvant therapy. METHODS: Review of medical records of patients with rectal cancer in HMSJ and HSJ Oncology Services since 2009 - cases submitted to neoadjuvant treatment with radio-chemotherapy. RESULTS: Of those 96 patients with this disease who underwent neoadjuvant therapy with radio-chemotherapy, 35 patients were eligible; complete tumor response was observed in 11 cases (31%), and 9 were submitted to surgical treatment. Comparing the leukocyte parameters between patients with complete response (CR) and incomplete response (IR) the following values were observed: total number of leukocytes (mean) CR 7390.9 × IR 7220.4 (p = 0.8); total lymphocytes CR 2103 × IR 1960.9 (p = 0.4); neutrophil/lymphocyte ratio CR 3.55 × IR 3.79 (p = 0.5). The mean radiotherapy dose was 49.1 Gy, with CR 47.3 × IR 50.0 (p = 0.06). CONCLUSION: It was not possible to demonstrate in this study a significant relationship between complete tumor response to neoadjuvant therapy with respect to blood leukocyte parameters analyzed. (AU)

INTRODUÇÃO: A abordagem radio-quimioterápica previa apresenta grande relevância no manuseio do câncer de reto, colaborando na preservação funcional do órgão, sendo realizada previamente à cirurgia. A resposta inflamatória tem papel importante neste tratamento. OBJETIVO: Consiste em correlacionar o número de linfócitos periféricos e a relação neutrófilos/linfócitos no sangue periférico com a resposta tumoral à terapia neo-adjuvante. MÉTODOS: Revisão de prontuários dos pacientes portadores de câncer retal dos serviços de Oncologia do HMSJ e HSJ, desde 2009, casos submetidos ao tratamento neo-adjuvante com radio-quimioterapia. RESULTADOS: Do total de 96 pacientes portadores desta enfermidade, submetidos à terapia neo-adjuvante com radio-quimioterapia foram elegíveis 35 pacientes, tendo sido observada resposta completa tumoral em 11 casos (31%), e nove foram submetidos ao tratamento cirúrgico. Na comparação dos parâmetros leucocitários entre os pacientes com resposta completa (RC) e resposta incompleta (RI) foram observados os seguintes valores: número total de leucócitos (média) RC 7390,9 × RI 7220,4 (p = 0,8); linfócitos totais RC 2103 × RI 1960,9 (p = 0,4); relação neutrófilo/linfócito RC 3,55 × RI 3,79 (p = 0,5). A dose radioterápica média foi de 49.1 Gy, sendo RC 47,3 × RI 50,0 (p = 0,06). CONCLUSÃO: Não foi possível demonstrar no presente estudo relação significativa entre resposta completa tumoral à terapia neo-adjuvante nos parâmetros analisados do perfil leucocitário. (AU)

Transcriptoma en mexicanos: metodología para analizar el perfil de expresión genética de gran escala en muestras simultáneas de tejido muscular, adiposo y linfocitos obtenidas en un mismo individuo / Transciptome among Mexicans: a large scale methodology to analyze the genetics expression profile of simultaneous samples in muscle, adipose tissue and lymphocytes obtained from the same individual

Objetivo: Describir la metodología de análisis de múltiples transcritos con técnicas de microarreglo en biopsias simultáneas de tejido muscular, adiposo y sangre en un mismo individuo, como parte de la estandarización del estudio GEMM (Genética de las Enfermedades Metabólicas en México). Material y métodos: Se incluyó a cuatro sujetos con índice de masa corporal (IMC) entre 20 y 41. Se registró estatura, talla y composición corporal. Se realizó biopsia muscular (vasto lateral), de tejido adiposo subcutáneo y muestra de sangre completa. El ARN total fue extraído de los tejidos y amplificado para análisis de microarreglos. Resultados: De 48 687 potenciales transcritos, 39.4% fue detectable en al menos uno de los tejidos. La expresión de leptina no fue detectable en linfocitos, débilmente expresada en músculo, alta expresión en el tejido adiposo y correlacionó con el IMC. El GLUT4 también ilustra la especificidad para el músculo sin verse afectado por el IMC. La concordancia en la expresión de transcritos fue 0.70 (p<0.001) para los tres tejidos. Conclusiones: Fue factible cuantificar simultáneamente la expresión genética de miles de transcritos, hubo concordancia en la expresión entre diferentes tejidos obtenidos en un mismo individuo, y confiabilidad del método al reproducir las relaciones biológicas esperadas. El estudio GEMM podrá analizar las correlaciones de los transcritos expresados dentro de un órgano y luego entre diferentes tejidos, y proveerá endofenotipos cuantitativos novedosos que proporcionarán un amplio panorama de información sobre las enfermedades metabólicas, incluyendo obesidad y diabetes tipo 2.

OBJECTIVE: We describe the methodology used to analyze multiple transcripts using microarray techniques in simultaneous biopsies of muscle, adipose tissue and lymphocytes obtained from the same individual as part of the standard protocol of the Genetics of Metabolic Diseases in Mexico: GEMM Family Study. METHODS: We recruited 4 healthy male subjects with BM1 20-41, who signed an informed consent letter. Subjects participated in a clinical examination that included anthropometric and body composition measurements, muscle biopsies (vastus lateralis) subcutaneous fat biopsies anda blood draw. All samples provided sufficient amplified RNA for microarray analysis. Total RNA was extracted from the biopsy samples and amplified for analysis. RESULTS: Of the 48,687 transcript targets queried, 39.4% were detectable in a least one of the studied tissues. Leptin was not detectable in lymphocytes, weakly expressed in muscle, but overexpressed and highly correlated with BMI in subcutaneous fat. Another example was GLUT4, which was detectable only in muscle and not correlated with BMI. Expression level concordance was 0.7 (p< 0.001) for the three tissues studied. CONCLUSIONS: We demonstrated the feasibility of carrying out simultaneous analysis of gene expression in multiple tissues, concordance of genetic expression in different tissues, and obtained confidence that this method corroborates the expected biological relationships among LEPand GLUT4. TheGEMM study will provide a broad and valuable overview on metabolic diseases, including obesity and type 2 diabetes.

Perfil da resposta Th1/Th2 no fluido gengival de pacientes portadores de doença inflamatória intestinal com periodontite crônica / Th cell profile in the gingival crevicular fluid from inflammatory bowel disease patients with chronic periodontitis

O objetivo dessa tese foi avaliar a expressão de citocinas Th1 (IL-12 e INFγ), citocinas Th2 (IL-4, IL-6 e IL-10) e das citocinas pró-inflamatórias IL-18, IL-1β e TNFα no fluido gengival de pacientes com periodontite crônica portadores da doença de Crohn (DC), de retocolite ulcerativa idiopática (RCUI) e em indivíduos saudáveis (o grupo controle, GC). Como objetivo secundário, avaliamos a função dos neutrófilos no fluido gengival desses pacientes através da mensuração das metaloproteinases da matriz -8, -9 (MMP-8 e MMP-9) e da atividade da elastase. Quinze pacientes com DC (idade média 38.2 ± 11.4 anos), 15 pacientes com RCUI (idade média 45.0 ± 10.5 anos) e 15 pacientes saudáveis (idade média 42.1 ± 7.8 anos) participaram desse estudo. Todos os dentes presentes, com exceção dos terceiros molares, foram examinados. Profundidade de bolsa (PB), nível de inserção clínica (NI), presença de placa e de sangramento a sondagem foram avaliados em seis sítios por dente. Em cada paciente, o fluido de 4 sítios com periodontite (PB ≥ 5 mm e NI ≥ 3mm) e de 4 sítios com gengivite (PB ≤ 3 mm e NI ≤ 1 mm) foram coletados através de pontas de papel absorvente pré-fabricadas. O sistema LUMINEX® foi utilizado na mensuração das IL-1β, IL-4, IL-6, IL-10, IL-12p70, TNFα, INFγ, MMP-8 e MMP-9. A IL-18 foi analisada através do ensaio ELISA e a atividade de elastase através de uma reação enzimática. O soro desses pacientes também foi analisado e o coeficiente de correlação de Pearson foi utilizado na análise da correlação entre as citocinas no soro e no fluido gengival. Nos sítios com gengivite, a quantidade total de IL-4 foi significativamente menor no grupo RCUI do que no grupo GC (p=0.016). Nos sítios com periodontite, a quantidade total de IL-4 foi significativamente menor no grupo DC do que no grupo GC (p=0.029)...

The aim of this thesis was to evaluate the expression of Th1 cytokines (IL-12 and INF-γ), Th2 cytokines (IL-4, IL-6 and IL-10) and the pro-inflammatory cytokines IL-18, IL-1β and TNF-α in the gingival crevicular fluid (GCF) from Crohn’s disease (CD) patients, ulcerative colitis (UC) patients and healthy individuals (control group, CG) who had chronic periodontitis. Besides, we measured elastase activity, matrix metalloproteinase -8 and -9 (MMP-8 and -9) to address the neutrophil function in the GCF. Fifteen CD patients (mean age 38.2 ± 11.4 years), 15 UC patients (mean age 45.0 ± 10.5 years) and 15 systemically healthy controls (mean age 42.1 ± 7.8 years) were enrolled in this study. All the present teeth, except for the third molars were examined. Probing pocket depth (PPD), clinical attachment loss (CAL), presence of plaque and presence of bleeding on probing were assessed in six sites per tooth. In every subject, GCF from 4 gingivitis sites (PPD ≤ 3mm and CAL ≤ 1mm) and from 4 periodontitis sites (PPD ≥ 5mm and CAL ≥ 3mm) were collected with filter strips. The data were reported as total amount and concentration. IL-1β, IL-4, IL-6, IL-10, IL-12p70, TNFα, INFγ, MMP-8 and MMP-9 were analyzed by the Luminex® analyzer. IL-18 was analyzed using a commercially available ELISA assay and the elastase activity by an enzymatic reaction. The serum was also analysed and the correlations between the cytokines in the GCF and in the serum were calculated by Pearson correlation analysis. In gingivitis sites, the total amount of IL-4 was significantly lower in the UC group than in the CG group (p=0.016). In periodontitis sites, the total amount of IL-4 was significantly lower in CD group than in the CG group (p=0.029). The total amount of IL-4 was lower in UC group than in CD group (p=0.077)...

Proliferating cell nuclear antigen (PCNA), p53 and MDM2 expression in Hodgkins disease

CONTEXT AND OBJECTIVE: Tumor cells in Hodgkins disease (HD) express cell proliferation markers that are evaluated according to the oncogenes involved or the expression of their proteins. Correlations between the protein expression grade and clinical data are now important for disease prognosis. DESIGN AND SETTING: This was a retrospective analysis on proliferating cell nuclear antigen (PCNA), p53 and MDM2 (murine double minute-2) expression using immunohistochemistry, on formalin-fixed, paraffin-embedded tissues from diagnostic biopsies on 51 patients with HD. The study was conducted at the Division of Hematology and Transfusion Medicine, Hospital São Paulo, Universidade Federal de São Paulo. METHODS: Antigen expression was evaluated as the proportions of positive Hodgkin and Reed-Sternberg (HRS) cells and reactive lymphocytes (L), which were compared using Spearman correlation coefficients. The Friedman test was used for comparisons between the markers. The Pearson test was used to investigate associations between marker expression and clinical and laboratory parameters, marrow involvement, complete remission (CR) and overall survival (OS) rates. RESULTS: There was overexpression of antigen proteins in HRS, in relation to L (p < 0.001). In HRS, MDM2 was higher than p53 and PCNA (p < 0.003), while the latter two were equivalent. In L, p53 was lower than MDM2 and PCNA (p < 0.001), while the latter two were equivalent. There was no relationship between protein expression and clinical and laboratory variables or outcome. CONCLUSIONS: PCNA, p53 and MDM2 are tumor markers for HD, but showed no clinical or prognostic significance in our analysis.

CONTEXTO E OBJETIVO: As células tumorais da doença de Hodgkin (HD) são positivas para marcadores de proliferação celular que são analisados por seus genes e respectivas proteínas. A correlação entre a expressão destas proteínas e os parâmetros clínico-laboratoriais são, no momento, de importância para o prognóstico da doença. TIPO DE ESTUDO E LOCAL: Estudo retrospectivo da expressão do antígeno de proliferação celular (PCNA) e da p53 e MDM2 em tecidos obtidos ao diagnóstico, fixados por formol, embebidos em parafina de 51 pacientes com HD. O trabalho foi realizado na Divisão de Hematologia e Transfusão, Hospital São Paulo, Universidade Federal de São Paulo. MÉTODOS: As expressões antigênicas foram analisadas através da proporção de células de Hodgkin e células de Reed Sternberg (HRS) e linfócitos reacionais (L) positivos. A intensidade de expressão de cada proteína foi comparada entre L e HRS através do coeficiente de Spearman. A comparação da PCNA, p53 e MDM2 em L e HRS se fez pelo teste de Fiedman. As correlações entre variáveis clínico-laboratoriais, comprometimento da medula óssea, taxas de sobrevida geral e remissão clínica com as proteínas em HRS se fizeram pelo coeficiente de Pearson. RESULTADOS: Houve superexpressão das três proteínas em células HRS comparadas aos L (p < 0,001). Nas células HRS, a MDM2 foi maior que a p53 e a PCNA (p < 0,003), que foram equivalentes. Nos L, a p53 foi menor que a MDM2 e a PCNA (p < 0,001), que foram equivalentes Não houve relação entre as expressões das proteínas com as variáveis clínico-laboratoriais e sobrevida. CONCLUSÕES: PCNA, p53 e MDM2 são marcadores tumorais na HD, porém não mostraram significado clínico-prognóstico em nossa análise.

Construction of a human Fab phage display library from antibody repertoires of osteosarcoma patients

Osteosarcoma is the commonest type of primary malignant bone tumor, frequently found in adolescents at sites of rapid bone growth. Despite current management protocols, up to half of the patients succumb to this disease. Moreover, there is no well-characterized molecular marker for diagnosis and prognosis. Since phage display methodology allows the selection of human antibody fragments with potential use in clinical applications, we applied this procedure to construct a recombinant Fab (antigen binding fragment) library from patients with osteosarcoma. We used peripheral blood lymphocyte total RNA from 11 osteosarcoma patients and cloned recombinant Fab representing the micro, gamma and kappa chain antibody repertoires of these individuals. The resulting library was cloned in the pComb3X vector and attained 1.45 x 10(8) different functional forms. BstO I fingerprinting and DNA sequencing analysis of randomly selected clones revealed the diversity of the library, demonstrating that Fab harbors Vkappa chains from subgroups I to V, biased towards the A27 fragment, as normally reported for the human repertoire. Analysis of the VH repertoire revealed that our library has a slight bias towards the VH4 family, instead of the usually reported VH3. This is the first description of a phage display library from osteosarcoma patients. We believe these human Fab fragments will provide a valuable tool for the study of this neoplasia and could also contribute to improvements in the diagnosis of this disease.

Análise imunocitoquímica do infiltrado inflamatório na miosite com corpo de inclusäo citoplasmática e em outras doenças neuromusculares com vacúolos marginados / Immunocytochemical analysis of the inflammatory infiltrate in inclusion body myositis and other neuromuscular disorders rimmed vacuoles

Encontramos 16 casos com vacúolos marginados entre 1400 biópsias musculares cujo diagnóstico final foi miosite com corpos de inclusao citoplasmática esporádica (MCIC) (4 casos), atrofia muscular espinhal juvenil (6 casos), miopatias distais (3 casos), distrofia das cinturas pélvica e escapular (2 casos) e neuropatia periférica (1 caso). Foram utilizados anticorpos monoclonais contra linfócitos T totais e subpopulaçoes (CD4+ e CD8+), linfócitos B, macrófagos, células exterminadoras naturais (NK), imunoglobulinas e porçao C3 do complemento. A análise foi quantitativa e de acordo com o local de acúmulo (interstício, intra-fibra e perivascular). Linfócitos CD8+ foram encontrados no interstício na maioria dos casos, sendo menos comuns dentro das fibras musculares e raros no espaço perivascular. Os casos de MCIC apresentaram maior número de linfócitos CD8+ se comparados às outras doenças. A proporçao de células CD8+/CD4+ foi maior na MCIC do que nas outras doenças. Existiam macrófagos em grande proporçao na MCIC, miopatias distais e em um dos casos de distrofia das cinturas pélvica e escapular. Células NK foram frequentes no interstício nos casos de MCIC e mais raras nas outras doenças. Houve maior depósito de imunoglobulinas e complemento nos casos de MCIC do que nas demais doenças. O grande número de células CD8+ e a relaçao CD8+/CD4+ podem auxiliar no diagnóstico diferencial da MCIC de outras doenças neuromusculares com vacúolos marginados.

Chromosome damage in underground coal miners: detection by conventional cytogenetic techniques and by submitting lymphocytes of unexposed individuals to plasma from at-risk groups

Estudos citogenéticos foram realizados em mineiros de carväo mineral de Criciúma (SC) e em controle fumantes e näo-fumantes selecionados por sexo e idade. Nas culturas de linfócitos da amostra de mineiros observamos uma diminuiçäo do índice mitótico e um excesso de fragmentos cromossômicos e em células da mucosa oral um excesso de micronucleos. Um ensaio alternativo para detecçäo de clastogênese em casos de exposiçäo ocupacional foi testado: os linfócitos de indivíduos näo expostos foram submetidos a uma mistura de plasmas de mineiros e também a uma mistura de plasmas de indivíduos näo-expostos, com a finalidade de servirem como controles pareados no experimento realizado. Os resultados revelaram que o teste é muito conveniente, uma vez que linfócitos dos mesmos indivíduos podem ser utilizados tanto como alvo como controle. Neste experimento detectamos um aumento de falhas cromatídicas, fragmentos e alteraçöes cromossômicas em geral nos linfócitos expostos a mistura de plasmas de mineiros. A mistura de plasmas dos controle näo induziu nenhuma alteraçäo significante, indicando que os resultados obtidos näo foram devidos ao efeito de adiçäo das misturas de plasmas per se.

Characterization of a lectin from goat peripheral blood lymphocytes.

A D-glucose specific lectin was isolated from goat peripheral blood lymphocytes by affinity chromatography on N-acetyl D-glucosamine agarose gel. The fluorescence intensity of 4 methyl umbelliferyl D-glucose was quenched to about 62% on addition of the lectin. This lectin gave a single band corresponding to 112 kDa in SDS-PAGE irrespective of treatment with 2-mercaptoethanol. The molecular weight and the Stoke's radius of the lectin in the native conditions were found to be 114 kDa and 4.54 nm, respectively, as determined by gel filtration on Sephacryl S 500 column. The lectin was found to be a glycoprotein with 5.6% of neutral hexose content and 5.5% of sialic acid. The lectin agglutinated trypsinized rabbit erythrocytes and human type A erythrocytes. The hemagglutinating activity was dependent on the presence of divalent cations like Mn2+ and Ca2+. Optimum pH, ionic strength and temperature for rebinding of lectin to acid treated Sephadex G200 were found to be 7.5, 0.16 and 30-37 degrees C, respectively.

Expression of HLA-DR antigen in large bowel carcinoma

One hundred large bowel carcinomas were studied immunohistochemically with regard to expression of HLA-DR antigen (DR). One or two sections from each tumor including surrounding normal mucosa were examined by a semiquantitative counting system for tumor cells and mucosal and stromal infiltrates of lymphocytes and mononuclear cells (MNCs) with DR expression and the results were applied Chi-square test. The rate of presence of DR positive (DR+) lymphocytes in lymphoid nodules and DR+ lymphocytes/ MNC in the adjacent mucosa and stroma in DR+ carcinoma (50%) was higher (P < 0.01) than in DR- carcinoma (21.9%). Thirty-six carcinomas (36%) were DR+. Three (75%) out of 4 DR+ poorly differentiated carcinomas and six (20%) out of 30 DR+ moderately differentiated carcinomas showed homogeneously strong DR+ expression. There was tendency for poorly differentiated carcinoma to be more homogeneous DR+ expression. According to Dukes' stage, four (80%) out of 5 carcinomas in Dukes' stage D were DR-. An increased infiltration of lymphocytes/MNCs into adjacent mucosa and stroma in large bowel carcinomas is possibly related with DR expression by carcinoma. From the results of this study, we postulated as follows: 1) DR+ tumor cells may act as antigen-presenting cells, 2) They may have an inhibitory effect for distant metastasis, 3) Poorly differentiated carcinoma expressed more DR+ homogeneously.

Basal and paracetamol-depleted glutathione from human lymphocytes: ethnic variability

El glutatión (GSH) se conjuga con una gran variedad de xenobióticos y sus metabolitos electrofílicos lo cual acelera su eliminación en la orina. De esta forma protege las células evitando la unión de estos intermediarios lábiles a moléculas vitales. Es posible que algunos individuos sean susceptibles a este efecto deletéreo en función de la concentración intracelular de GSH. En el presente estudio se pretende determinar la magnitud y naturaleza de la variación interindividual, post-stress oxidativo con paracetamol, del contenido de glutatión en linfocitos humanos. Se midió el GSH intracelular luego de una hora de incubación de los linfocitos en un edio de cultivo adicionado con microsomas de hígado de rata y NADPH A 37ºC. Se midieron en paralelo dos lotes de linfocitos cada vez, uno sin agregar paracetamol al medio (basal) y otro agregando paracetamol al medio (deplecionado por paracetamol). Se estudiaron 90 individuos adultos Caucásicos en los cuales la concentración basal media de GSH linfocitario (22,3µmol/10**7 células) fue mayor que un grupo comparable de 20 Chinos (19,1mol/10**7 células y de 20 individuos Afroamericanos (17,9µmol/10 7 celulas). El GSH delos linfocitos sometidos a paracetamol disminuyó en la misma proporción en cada grupo, de manera que las mismas diferencias vistas en condiciones basales persisten post stress oxidativo. El análisis de los valores de cada población mediante la prueba del probit reveló una distribución unimodal. En un grupo de individuos caucásicos con síndrome de Down la concentración de GSH basal (29,7 µmol/10**7 células) fue más alta que en el resto de los grupos estudiados

Lymphocyte-bearing ferritin in beta-thalassemia/Hb E.

Flow cytometric analysis of lymphocyte subsets and lymphocyte surface ferritin shows no significant difference in the number of total T-cells, B cells, NK cells, helper T-cells (CD 4), suppressor T-cells (CD 8) and CD 4/CD 8 ratio among normal subjects (n = 11) and Hb E trait (n = 6), beta-thalassemia (beta-thal) trait (n = 5), neither in normal and nonsplenectomized patients with beta-thal/Hb E (n = 10) except B cells and CD 4. There is a significant reduction in lymphocytes surface spleen-type and heart-type ferritin in patients with beta-thal/Hb E when compared to normal subjects. No difference can be seen among patients with beta-thal/Hb E, beta-thal trait and Hb E trait. This low percentage of lymphocyte-bearing ferritin suggests a negative relationship between ferritin on the cells' surface and high circulating ferritin normally associated with thalassemic patients.