Peritoneal fluid collection in healthy cattle and evaluation of changes in cell morphology during storage in refrigerated and non-refrigerated samples / Colheita do fluido peritoneal de bovinos sadios e verificação das alterações na morfologia celular quando mantido refrigerado ou à temperatura ambiente

This study aimed to evaluate the appropriate sites of abdominocentesis for peritoneal fluid collection in cattle and to investigate the time of cell viability in vitro, comparing three methods of sample conservation. Twenty-one healthy cattle (19 females and 2 males) were subjected to a laparocentesis procedure to obtain peritoneal fluid, with punctures in three defined sites: left cranial, right cranial, and right caudal. The total peritoneal fluid collected was divided into three aliquots and maintained under three preservation conditions: room temperature (26°C), refrigeration (4°C), and room temperature (26°C) with the addition of 1µL of 10% formaldehyde per 1mL of peritoneal fluid. The peritoneal fluid analysis performed immediately after collection consisted of: physical examination (color, appearance, volume, and specific gravity), biochemical measures (pH, total protein, fibrinogen, creatinine, and glucose), and cellularity (total and differential counts). The determination of proteins and the examination of cells were repeated in each separate aliquot at two, four, six, and eight hours after harvest. Data were analyzed through repeated measures ANOVA or Friedman test. The harvest was productive in 67% of cattle. The left cranial and the right cranial puncture sites were the most appropriate. Peritoneal fluid analyzed after collection, the total protein concentration ranged from 1.4 to 3.6g/dL, and number of leukocytes ranged from 54 to 1,322 cells/µL; 60 to 95% of leukocytes were lymphocytes. The protein concentration decreased, but the absolute values of leukocytes, lymphocytes, and segmented neutrophils did not change up to eight hours after collection, independent of the maintenance method. Cell lysis was delayed by cooling, and the addition of formaldehyde did not help preserve the integrity of cellular morphology. Laparocentesis is a safe and secure procedure in cattle and maybe more productive when performed in specific sites on the left or right sides of the cranial abdominal wall. Peritoneal fluid samples may be analyzed with reliable results for up to eight hours after collection when kept refrigerated and for up to six hours when kept at room temperature.(AU)

O estudo teve como objetivo avaliar os locais adequados de laparocentese para a colheita de fluido peritoneal de bovinos e estabelecer o tempo de viabilidade celular in vitro, comparando três métodos de conservação. Vinte e um bovinos hígidos (19 fêmeas e 2 machos) foram submetidos ao procedimento de laparocentese para obtenção de fluido peritoneal, com punção em três pontos definidos: cranial esquerdo, cranial direito e caudal direito. O volume total do líquido peritoneal foi dividido em três alíquotas mantidas sob três métodos de conservação: temperatura ambiente (26°C); refrigeração (4°C); e temperatura ambiente (26°C) com adição de 1µL de formol 10% para cada 1mL de líquido peritonial. A análise do líquido peritoneal realizada imediatamente após sua obtenção consistiu em: exames físico (cor, aspecto, volume e densidade); bioquímicos (pH, proteína total, fibrinogênio, creatinina e glicose); e da celularidade (contagens total e diferencial). A determinação de proteínas e o exame da celularidade foram repetidos, em cada alíquota separada, as duas, quatro, seis e oito horas após a colheita. Análise de variâncias de medidas repetidas ou teste de Friedman foram empregados para avaliação ao longo do tempo. A colheita foi produtiva em 67% dos bovinos e os locais de punção craniais esquerdo e direito foram os mais adequados. A concentração de proteína total variou de 1,4 a 3,6g/dL e o número de leucócitos de 54 a 1.322 células/µL, com predomínio de linfócitos (60 a 95% das células) no fluido peritoneal analisado logo após a colheita. A concentração de proteínas diminuiu, mas os valores absolutos de leucócitos, de linfócitos e de neutrófilos segmentados não se modificaram até oito horas após a colheita, independente do método de manutenção das amostras. A lise celular foi retardada pela refrigeração e a adição de formol não contribuiu para preservar a integridade da morfologia celular. A laparocentese é um procedimento seguro e de execução fácil em bovinos sendo mais produtiva quando realizada em locais específicos à esquerda ou à direita craniais da parede abdominal. Amostras de fluido peritoneal podem ser analisadas com resultados confiáveis quando mantidas refrigeradas por até oito horas após a colheita e quando mantidas à temperatura ambiente por até seis horas.(AU)

A Case of Residual Non-Small Cell Lung Cancer Cells Coexisting With Newly Developed Small Cell Lung Cancer Cells in Ascitic Fluid after Chemotherapy for Non-Small Cell Lung Cancer

No abstract available.

Cytological Study of Effusions.

Background: Effusion fluid analysis plays an important role in clinical medicine. Clinicians rely on the reports of effusion fluids and use them as complement to their clinical assessment for the diagnosis and management. Aim: To study the incidence of neoplastic and non neoplastic effusions. Objectives: i) To study the gross and microscopic features of effusions; ii) To study the pattern of effusions in various neoplastic and non neoplastic conditions. Material and Method: 550 specimens of pleural, peritoneal and pericardial fluid were studied. Fluid samples were centrifuged for five minutes at 2000 rpm and smears prepared from deposit were stained by Haematoxylin and Eosin (H and E), Giemsa and Papanicolaou stains (Pap). Result: Out of 550, 315 were pleural effusions, 234 peritoneal and one was pericardial. Out of total 315 cases of pleural effusions, 297 were non neoplastic and 18 were neoplastic effusion. Out of total 234 peritoneal effusions 214 were non neoplastic and 20 neoplatic. Commonest malignancy in pleural and peritoneal fluid was adenocarcinoma. Conclusion: Pleural effusion was the commonest fluid in this study. Exudates were predominant in pleural effusion and transudates were predominant in peritoneal effusion. Common causes of exudates in pleural effusion were tuberculosis (TB), pneumonia and malignancy. Common causes of transudates in peritoneal effusion were liver cirrhosis and congestive cardiac failure (CCF). Adenocarcinoma was the commonest malignancy in both pleural and peritoneal effusion (30 cases).

Peritoneal fluid changes in horses subjected to small colon distension / Alterações no líquido peritoneal de equinos submetidos a distensão do cólon menor

Intestinal devitalization in cases of small colon obstruction may be difficult to detect based only in clinical signs. The purpose was to serially evaluate blood and peritoneal fluid of horses subjected to small colon distension. Seventeen adult horses were allotted in three groups. In the small colon-distended group (DG, n=7) a surgically-implanted latex balloon was inflated to promote intraluminal small colon distension. In the shamoperated group (SG, n=5), the balloon was implanted but not inflated, and no surgery was done in the control group (CG, n=5). Blood and peritoneal fluid were sampled before and after (6 samples with a 30-minute interval) intestinal obstruction for cytological and biochemical analyses. No significant changes in clinical signs occurred within groups or across time during the experimental period. There were no statistical differences among SG and SG groups in hematologic and blood chemistry variables. Although total protein concentration and lactate dehydrogenase (LDH) activity in peritoneal fluid remained most of the time within reference values during the experimental period in all groups, increases from baseline values were detected in SG and DG groups. Such increases occurred earlier, progressively and with greater magnitude in the DG when compared with the SG (P<0.05). Increases from baselines values were also observed in total nucleated cells and neutrophils counts in the DG (P<0.05). In conclusion, distension of the equine small colon induced progressive subtle increases in total protein and LDH concentrations in the peritoneal fluid during the first hours. Serial evaluation of these variables in peritoneal fluid may be useful for early detection of intestinal devitalization in clinical cases of equine small colon obstruction.

A desvitalização do cólon menor em equinos pode ser difícil de ser detectada baseando-se apenas em sinais clínicos. O objetivo foi realizar uma avaliação seriada do líquido peritoneal de equinos submetidos à distensão do cólon menor. Dezessete cavalos adultos foram divididos aleatoriamente em três grupos. No grupo distendido (DG, n=7) um balão implantado cirurgicamente foi inflado para promover distensão do cólon menor. No grupo instrumentado (SG, n=5) o balão foi implantado, mas sem promover distensão e no grupo controle (CG, n=5) não houve anestesia ou cirurgia. Sangue e fluido peritoneal foram colhidos antes e durante 180 minutos após a cirurgia para análises citológicas e bioquímicas. Nenhuma interação significativa ocorreu entre grupos e tempos nas variáveis clínicas e hematológicas. Apesar dos valores de proteínas totais e da atividade da lactato desidrogenase (LDH) permanecerem dentro da normalidade durante quase todo o experimento, aumentos em relação aos valores basais ocorreram nos grupos SG e DG. Contudo, tais aumentos foram precoces, progressivos e em maior magnitude em DG quando comparados ao SG, mostrando que a distensão promoveu alterações significativas nessas variáveis (P<0.05). Aumentos em relação aos valores basais também ocorreram nas contagens de células totais nucleadas e neutrófilos (P<0.05). Em conclusão, a distensão experimental do cólon menor promove, nas primeiras horas, alterações subliminares progressivas nas concentrações de proteínas totais e na atividade de LDH no líquido peritoneal. Os resultados indicam que a avaliação seriada do liquido peritoneal pode ser útil para detectar desvitalização intestinal em casos clínicos de obstrução do cólon menor equino.

Comparison of role of serum- ascites albumin gradient and ascitic fluid total protein in liver cirrhosis patients

To compare the diagnostic sensitivity of serum/ ascites albumin gradient and ascitic fluid total protein in liver cirrhosis patients, using ultrasonography as gold standard. Validation Study. Department of Chemical Pathology and Endocrinology, Armed Forces Institute of Pathology, Rawalpindi and Department of Radiology CMH/ MH Rawalpindi from 15 Jul 2007 to 15 May 2008. Seventy three patients of liver cirrhosis were enrolled in the study by non-probability convenience sampling. Liver cirrhosis was confirmed on ultrasound abdomen. Ascitic fluid and 3 ml of blood were obtained simultaneously for analysis of serum albumin, ascitic fluid albumin and total proteins. Sensitivity of serum ascitic albumen gradient [SAAG] and serum ascitic fluid total protein [AFTP] was calculated by comparing with liver ultrasonographic findings [gold standard]. Among 73 patients, 52 [71%] were males and 21 [29%] females. Mean ages was 57 years. Age range was 30-80 years. It was observed that sensitivity of SAAG in liver cirrhosis was 97% and that of AFTP was 53% only. Diagnostic sensitivity of SAAG in liver cirrhosis is significantly higher than AFTP in workup of ascites related to portal hypertension

Value of cytokeratins and calretinin in the differentiation between reactive mesothelial cells and malignant epithelial cells in cell block preparation

Malignant epithelial cells can be difficult to distinguish from reactive benign mesothelial cells in cytological smears prepared from body fluid effusions. Calretinin and cytokeratin 5/6 are valuable markers in the immunohistochemical distinction between malignant mesothelioma and adenocarcinoma in tissue sections. However, there is limited and conflicting data regarding their utility in the differential diagnosis between reactive benign mesothelial cells and malignant cells in cytologic material. The aim of the current work is to evaluate the efficiency of IHC staining of cairetinin, CK5/6, CK7, CK20 and TTF-1 in the distinction between reactive mesothelial cells and metastatic adenocarcinoma and the prediction of origin of primary carcinoma in cell blocks prepared from ascitic and pleural effusions. Formalin-fixed paraffin-embedded cell block preparations of 20 cytologic fluids were stained immunohistochemically for monoclonal antibodies to cairetinin, CK5/6, CK7, CK20, TTF-1 and CD68. Mesothelial cells showed positive immunoreactivity to calretinin and CK5/6 [100%], and CD68 [30%] while they were totally negative to CK7, CK20, and TTF-1. In contrast, malignant epithelial cells [in all studied cell block preparations] were negative to cairetinin, CK5/6 and CD68 while 50% were immunopositive to CK7, 34% to CK 20, and 15% to TTF-1. Cell block preparation is an excellent method for immunohistochemical staining of cytologic specimens. Calretinin and CK5/6 are valuable markers in the identification of reactive mesothelial cells. The use of an immunohistochemical panel of antibodies to calretinin, CK5/6, CK7, CK20, and TTF-1 is useful in the differential diagnosis between reactive mesothelial cells and metastatic adenocarcinoma in cytologic cell block preparations

Características celulares e bioquímicas do líquido peritoneal de equinos submetidos à obstrução experimental do duodeno, íleo e cólon maior / Cellular and biochemical characteristics of peritoneal fluid of equines submitted to experimental obstruction of duodenum, ileum, and large colon

Estudaram-se as características macroscópicas, bioquímicas e citológicas do líquido peritoneal de equinos submetidos a um modelo experimental de obstrução intestinal em 24 animais, distribuídos em grupos: de controle instrumentado (GI), de obstrução do duodeno (GII), de obstrução do íleo (GIII) e de obstrução do cólon maior (GIV). As amostras de líquido peritoneal foram colhidas antes da cirurgia (T0), durante as obstruções (T60i-T180i) e após as desobstruções (T60ri-T7º ). Durante o período obstrutivo, não foram observadas alterações significativas nos parâmetros bioquímicos e citológicos avaliados no líquido peritoneal dos animais. Após as desobstruções, apenas os animais de GII e GIII apresentaram resposta inflamatória mais intensa, caracterizada por maior contagem global e diferencial de leucócitos e dos valores de proteína total, fibrinogênio, lactato e fósforo inorgânico. As alterações laboratoriais não foram associadas a sinais clínicos indicativos da presença de processo inflamatório abdominal, demonstrando que os resultados da análise do líquido peritoneal, apesar de auxiliarem no acompanhamento da evolução do processo de cura, não devem ser utilizados isoladamente na elaboração do diagnóstico e do prognóstico de complicações no pós-operatório de equinos com cólica.

The macroscopic, biochemical, and cytological characteristics of peritoneal fluid of equines submitted to intestinal obstruction using an experimental model were evaluated. Twenty-four animals were distributed in four groups: instrumented control (GI), duodenum obstruction (GII), ileum obstruction (GIII), and large colon obstruction (GIV). Peritoneal fluid samples were collected before the surgery (T0), during the obstruction (T60i-T180i), and after unblocking procedures (T60ri-T7º ). During obstructive period, significant alterations were not observed in biochemical and cytological examination of peritoneal fluid of all animals. After unblocking procedure, animals from GII and GIII presented intense inflammatory response characterized by higher global and differential leukocytes counts, as well as in fibrinogen and total protein concentrations; lactate and inorganic phosphorus concentrations in peritoneal fluid were also increased. The laboratory alterations were not associated with clinical signs, indicative of the abdominal inflammatory process presence. Results showed that analyses of peritoneal fluids can be used as support in the evolution of healing process. However, they can not be isolated used for a diagnosis and prognosis of equine postoperative complications.

Utility of AgNor, immunocytochemistry for CEA and tumor markers for diagnosis in serous fluids

Objective: The evaluation of serous fluids stained by morphological methods lacks, in many cases, the necessary accuracy to obtain the correct diagnostics. The objective of this work was to establish the value of complementary tools for the improvement of diagnosis in serous effusions. Methods: Fifty-six serous effusions were processed for morphological staining, immunocytochemistry of carcinoembryonic antigen (CEA), AgNOR counting and electrochemiluminescense immunoassay for tumor markers (TM): CEA, Ca125 and CYFRA 21-1. TM assays were also performed in sera from the same patients. The Sensitivity (Se) and Specificity (Sp) were evaluated for all the methods. Results: Cytology: Se 73 per cent, Sp 100 per cent, CEA by immunocytochemistry: Se 96 per cent, Sp 75 per cent, AgNOR:Se 86 per cent, Sp100 per cent, TM: a) in fluids: CEA, Ca125 and CYFRA 21-1, Se: 29 per cent, 66 per cent and 64 per cent respectively and Sp: 100 por cento, 87 por cento and 100 por cento respectively. b) in sera: CEA, Ca125 and CYFRA 21-1: Se: 27 per cent, 77 per cent and 47 per cent respectively and Sp: 100 per cent, 25 per cent and 75 per cent respectively. CEA (in cells) + TM (fluids): Se 100 per cent and Sp 75 per cent AgNOR + TM (fluids): Se 95 per cent and Sp 87 per cent TM Panel (CEA+Ca125+CYFRA 21-1): a) in fluids: Se 81 per cent and Sp 87 per cent b) in sera: Se 86 per cent and Sp 12 per cent Conclusion: AgNOR assay and immunocytochemistry for CEA were useful as complementary tools in the diagnosis using effusions, raising the Sensitivity of the Cytology from 73 per cent to 86 per cent and 96 per cent respectively. Sensitivity increased with the assays for a panel of TM in fluids, but the high costof these methods does not justify their use for non-conclusive smears.

Cytology of body fluids from different sites: an approach for early diagnosis of malignancy.

Malignant effusions are a common presenting sign of malignancy and reflect dissemination. A retrospective study of all fluid samples accessioned at the Department of Pathology, TUTH from April 2000 to October 2002 were done. Over the study period, a total of 584 specimens were examined- 324 peritoneal fluid, 224 pleural fluid, 19 pericardial fluid, 9 knee joint effusion and 8 Cerebro-Spinal Fluid (CSF). One hundred and nine (18.66%) out of 584 cases were found to have malignancy, 57 were male and 52 were female. The age group of the adult male ranged from 42-78 years and female ranged from 43-62 years. Three patients were children with age ranging from 8-11 years. Adenocarcinoma was the commonest that comprised 89%, followed by Non Hodgkin's lymphoma 6.5% squamous cell carcinoma 2.7% and small cell carcinoma comprised 1.8 %. Exfoliative cytology is cheap, rapid and highly effective tool for the evaluation of body fluid and should be advised in all effusion cases.

Evaluation of reagent strips for ascitic fluid leukocyte determination: is it a possible alternative for spontaneous bacterial peritonitis rapid diagnosis?

In order to evaluate the accuracy of a urine reagent dipstick (Multistix 10SG®) to determine ascitic fluid leukocyte count, we prospectively studied 106 cirrhotic patients from April 2003 to December 2004, in two different centers (Federal University of São Paulo - UNIFESP-EPM and Federal University of Juiz de Fora - HU-UFJF) for the rapid bedside diagnosis of spontaneous bacterial peritonitis. The mean age 54 ± 12 years, there was a predominance of males (eighty-two patients, 77 percent), and alcohol was the most frequent etiology (43 percent). Forty-four percent of patients were classified as Child B and fifty-one as Child C (51 percent). Abdominal paracentesis was performed both in outpatient and inpatient settings and the Multistix 10SG® was tested. Eleven cases of spontaneous bacterial peritonitis were identified by means of polymorphonuclear count. If we considered the positive Multistix 10SG® result of 3 or more, the sensitivity, specificity, positive and negative predictive value were respectively 71 percent, 99 percent, 91 percent and 98 percent. With a positive reagent strip result taken as grade 2 (traces) or more, sensitivity was 86 percent and specificity was 96 percent with positive and negative predictive values of 60 percent and 99 percent, respectively. Diagnostic accuracy was 95 percent. We concluded that the use of a urine reagent dipstick (Multistix 10SG®) could be considered a quick, easy and cheap method for ascitic fluid cellularity determination in SBP diagnosis.

Diagnostic approach of atypical cells in effusion cytology using computerized image analysis

Cytology is one of the important diagnostic tests done on effusion fluid. It can detect malignant cells in up to 60% of malignant cases. The most important benign cell present in these effusions is the mesothelial cell. Mesothelial atypia can be striking and may simulate metastatic carcinoma. Many clinical conditions may produce such a reactive atypical cells as in anemia, SLE, liver cirrhosis and many other conditions. Recently many studies showed the value of computerized image analysis in differentiating atypical cells from malignant adenocarcinoma cells in effusion smears. Other studies support the reliability of the quantitative analysis and morphometric features and proved that they are objective prognostic indices. Sixty three cases of pleural and peritoneal smears, previously reported as benign [19] cases, malignant [21] cases or atypical [23] cases, were retrieved from the files. In each of these smears; nuclear area, perimeter, and roundness coefficient of 80-100 cell were determined at x400 magnification by the use of image analysis system. Statistical analysis was performed using analysis of variance and Tukey's HSD test. The mean values of nuclear roundness, nuclear perimeter and nuclear area vary between the three groups [benign, atypical and malignant cells] by using analysis of variance [p > 0.01]. The value of nuclear roundness, perimeter and area did not differ significantly between benign and atypical cells [Tukey's test: p<0.01]. On the other hand, the value of nuclear roundness, perimeter and area showed a significant difference between malignant and atypical cells [Tukey's test: p> 0.01]. In conclusion, our data suggest that cytomorphometry performed on effusion smear cells may provide important information for the differentiation of atypical cells from malignant cells, in which the values of atypical cells are closer to those of benign cells during the examination of pleural and peritoneal smears by the use of image analysis system

Study of effusion cytology in patients with simultaneous malignancy and ascites.

OBJECTIVE: To evaluate sensitivity of effusion cytology in detecting malignancy. MATERIALS AND METHODS: Effusion cytology was studied from 37 malignancy associated and 28 non malignancy associated ascitic fluid samples. RESULTS: Out of 65 cases, 44 (67.7%) effusions were reported negative, 15 (23.1%) were positive and 6 (9.2%) were suspicious for malignancy. Thus total 21 effusions (32.3%) were tumour cell positive. All 21 (100%) were true positive, none (0%) was false positive, 28 (63.6%) were true negative and 16 (36.4%) were false negative. Thus ascitic fluid cytology had sensitivity of 56.7% and specificity of 100%. Predictive value of positive test and negative test was 100% and 63.6% respectively. Stomach was the most common primary site of malignancy associated with ascites (11/37 i.e. 29.7%) where as adenocarcinoma was the most common type of malignancy (11/15 i.e.73.3%) in ascitic fluid cytology. CONCLUSION: Ascitic fluid cytology is a simple and useful procedure with sensitivity of 56.7% and should be routinely requested.

Pelvic endometriosis a Rare Cause of massive hemorrhagic Ascites

Endometriosis is defined as the presence of endometrial tissue outside the uterine cavity. It generally involves the peritoneum, ovaries and rectovaginal septum. Recurrent hemorrhagic ascites secondary to endometriosis is an unusual occurrence, 42 cases have been reported worldwide since 1954. It is more commonly seen in black nulliparous females. We report a case to draw attention to this condition as a potential cause for massive hemorrhagic ascites

Patterns of argyrophilic nucleolar organiser regions in pleural and peritoneal effusions

To evaluate AgNOR size and dispersion as alternate methods to AgNOR counts in order to differentiate malignant from non-malignant effusions. Comparative study. Department of Pathology, Postgraduate Medical Institute, Lahore, from January 2003 to June 2004. A total of 240 samples of pleural and peritoneal effusions were centrifuged, deposits smeared on slides and stained with H and E and AgNOR stain. The diagnosis of malignancy or otherwise was made on H and E staining. AgNOR counts, variation in size and dispersion of AgNOR dots in smears were graded and compared in malignant and non-malignant effusions. Mean AgNOR counts of 11.47 +/- 3.60 and 11.04 +/- 3.89 in malignant pleural and peritoneal effusions, respectively, were significantly [p<0.0001] greater as compared with counts of 3.36 +/- 0.69 and 3.35 +/- 0.66 in non-malignant effusions. AgNOR size and dispersion were of higher grade in significantly greater proportion of malignant as compared with non-malignant effusions [p<0.0001]. Typing of AgNOR size and dispersion was found to be an easy and reproducible alternative to traditional AgNOR counts for differentiating malignant from non-malignant effusions. These parameters should be correlated with already established but expensive techniques of AgNOR area and size imaging by electron microscopy and flow cytometry, as an economical alternative

Comparative study between the concentration of interleukin-6 in peritoneal fluid and in serum of patients with endometriosis

Our purpose was to compare the level of interleukin- 6 in peritoneal fluid with that in serum in patients with and without endometriosis undergoing laparoscopic procedures and to set a value for Interleukin 6 as a marker for endometriosis in the serum and peritoneal fluid of patients diagnosed laparoscopically to have endometriosis and correlate it with the stage of the disease. The present study was conducted at the period starting from March 2002 to October 2003 in the laparoscopy unit, Ain Shams University Maternity hospital. Fifty patients underwent laparoscopy were recruited in this case control cross-sectional study. These patients were divided into two groups. Group I included thirty patients diagnosed to have endometriotic lesions by means of laparoscopy and histopathologic confirmation. Group II included twenty patients diagnosed laparoscopically to have no visible gynecological lesions [e.g. women undergoing laparoscopic tubal ligation]. These patients were subjected to full history taking with special regard to symptoms suggestive of endometriosis, full general, abdominal and local examination for signs of endometriosis, laparoscopic visualization and staging of endometriosis according to the criteria of the revised American Fertility Society classification system, testing for serum and peritoneal fluid Interleukin-6 using the BioSource International ELISA. [Enzyme linked immunosorbent assay], and histopathologic confirmation of peritoneal endometriotic lesions. Interleukin- 6 was significantly higher among female patients diagnosed to have endometriosis than in control patients both in serum and peritoneal fluid. Also, our study revealed a highly significant direct correlation between the level of serum and peritoneal fluid IL-6 among women with endometriosis. A significant direct correlation between the level of IL-6 with the grade of endometriosis among cases was found both in serum and peritoneal fluid IL-6. However, no significant correlation was found between the grade of endometriosis and pain among the cases group. Increased peritoneal fluid levels of interleukin-6 in patients with active red endometriosis may relate to endometriosis-associated infertility and to the pathogenesis of endometriosis

Study of some biochemical assays in serum and ascitic fluid for differentiation between cirrhotic and malignant ascites

The clinical diagnosis of ascites is an easy task but the etiological diagnosis of the type of ascites is occasionally a different problem. Many studies have investigated the possibility of using a single marker to detect cancer in ascitic patients; but no analysis of ascitic fluid including cytology has been shown to be specific and sufficiently sensitive for either hepatic or extrahepatic cancer. To evaluate the role of various biochemical parameters including cholesterol, LDH, total protein, albumin and glucose for the differentiation of cirrhotic ascites from malignancy related ascites, fifty ascitic patients were classified into three main groups: Group I: included 20 patients with cirrhotic ascites. Group II: included 15 ascitic patients with hepatocellular carcinoma on top of liver cirrhosis. Group III: included 15 ascitic patients with extrahepatic malignancy without liver cirrhosis. All patients were subjected to: clinical history and examination, various imaging, routine laboratory investigations, histopathological examination, cytological examination of ascitic fluid, and biochemical analysis of both serum and ascitic fluid. We found that, although cytological examination of ascitic fluid is specific, it is less sensitive than ascitic fluid biochemical analysis in determining the cause of ascites. The accuracy of ascitic fluid cholesterol LDH, SAAG, A/S LDH ratio, A/S cholesterol ratio, A/S protein ratio, AFTp and ascitic/blood glucose ratio in discrimination of exudative malignant ascites from transudative cirrhotic ascites are [91%, 91%, 91%, 86%, 83%, 83%, 80% and 71%] respectively. Ascitic fluid biochemical analysis may be useful than cytological examination in follow-up of cirrhotic ascitic patients aiming to early detection of complication

Differential diagnosis of ascites: biochemical basis

This study was conducted to detect and assess the value of different biochemical parameters in the serum and ascitic fluid in differential diagnosis of ascites. The study included 50 patients with ascites [26 with liver cirrhosis, 14 malignant and 10 tuberculous]. Serum and ascitic fluid determination of total proteins, albumin, cholesterol, lactic dehydrogenase [LDH], Nitric Oxide [NO], ferritin and glucose. The mean value of total serum proteins and albumin was low in cirrhotic patients than in malignant and tuberculous ones but no significant difference was found between these groups. Mean values of ascitic fluid serum proteins and albumin was low in cirrhotic patients with a highly significant difference between cirrhotic and both malignant and tuberculous [P < 0.0001], but no significant difference between malignant and tuberculous ascites was found. All cirrhotic patients had SAAG >/= 1.1 [100%] versus 3 [21.5%] of malignant and 5 [50%] of tuberculous group. No significant difference between the mean value of serum cholesterol between the three groups but the mean value of ascitic fluid cholesterol was found [26.8 +/- 5.2, 92.3 +/- 18.8 and 59.5 +/- 9.7 mg/dL] in cirrhotic, malignant and tuberculous respectively. A highly significant difference in mean value of ascitic fluid cholesteral between cirrhotics and both malignant and tuberculous groups [P < 0.0001] was present and a significant difference between malignant and tuberculous [P < 0.05] levels was also found. There was no significant difference between the mean value of serum LDH between the three groups but ascitic fluid LDH was very high in malignant group than both cirrhotic and tuberculous [58.7 +/- 12.4] versus 33.2 +/- 11 and 30.2 +/- 5.6 IU/L]. A highly significant statistical difference in the mean value of ascitic fluid LDH between malignant and each of cirrhotic and tuberculous levels [P < 0.001]. Both NO and ferritin were slightly high in ascitic fluid of cirrhotic and malignant patients but no statistical difference was found between groups either in serum or ascitic fluid. There was no statistical differences between the mean levels of serum glucose in all groups but a significant [P < 0.05] lower level was found in ascitic fluid of the tuberculous group compared with the other two groups. Conclusions: From this study we can conclude that diagnostic paracentesis is a useful procedure. The practice of ordering a battery of tests on every ascitic fluid specimen should be abandoned. Rather, an algorithm approach should be adopted in which the result of initial analysis guide us to further relevant tests that help in arriving at the etiology of ascites. From our result we can apply the following: 1- Ascitic fluid albumin, ascitic fluid/serum albumin, total proteins and SAAG are the best parameters for diagnosis of sterile cirrhotic ascites. 2- Ascitic fluid cholesterol and LDH in combination in addition to high ascitic fluid proteins are useful for diagnosis of malignant ascites. 3- Ascitic fluid glucose, ascitic fluid/serum glucose ratio and high ascitic fluid total proteins were preferred for diagnosing tuberculous ascites

T cell phenotype and intracellular IFN-gamma production in peritoneal exudate cells and gut intraepithelial lymphocytes during acute Toxoplasma gondii infection in mice

Although there are many reports on the splenic (systemic) T cell response after Toxoplasma gondii infection, little information is available regarding the local T cell responses of peritoneal exudate cells (PEC) and gut intraepithelial lymphocytes (IEL) following peroral infection with bradyzoites. Mice were infected with 40 cysts of the 76K strain of T. gondii, and then sacrificed at days 0, 1, 4, 7 and 10 postinfection (PI). The cellular composition and T cell responses of PEC and IEL were analyzed. The total number of PEC and IEL per mouse increased after infection, but the ratio of increase was higher in IEL. Lymphocytes were the major component of both PEC and IEL. The relative percentages of PEC macrophages and neutrophils/eosinophils increased significantly at day 1 and 4 PI, whereas those of IEL did not change significantly. The percentage of PEC NK1.1 and gamma delta T cells peaked at day 4 PI (p < 0.0001), and CD4 and CD8 alpha T cells increased continuously after infection. The percentages of IEL CD8 alpha and gamma delta T cells decreased slightly at first, and then increased. CD4 and NK1.1 T cells of IEL did not change significantly after infection. IFN-gamma-producing PEC NK1.1 T cells increased significantly from day 1 PI, but the other T cell subsets produced IFN-gamma abundantly thereafter. The proportion of IEL IFN-gamma-producing CD8 alpha and gamma delta T cells increased significantly after infection, while IEL NK1.1 T cells had similar IFN-gamma production patterns. Taken together, CD4 T cells were the major phenotype and the important IFN-gamma-producing T cell subsets in PEC after oral infection with T. gondii, whereas CD8 alpha T cells had these roles in IEL. These results suggest that PEC and IEL comprise different cell differentials and T cell responses, and according to infection route these factors may contribute to the different cellular immune responses.

Inducción de la síntesis de óxido nítrico en células mononucleares en cultivo utilizando líquido peritoneal de mujeres con endometriosis, en relación al porcentaje de linfocitos T y células NK identificado en dicho ambiente / Induction of nitric oxide synthesis in mononuclear cells with peritoneal fluid relatin to T-lymphocyte and NK cells in patients with endometriosis

A pesar de que la endometriosis representa un problema de salud reproductiva de gran importancia por ser una de las afecciones ginecológicas benignas más comunes, aún se desconoce su etiopatogenia. La hipótesis más aceptada es la de John Sampson la cual propone que células y tejido endometrial provenientes del fujo menstrual durante la descamación uterina llegan al peritoneo a través de tubas por contraflujo bajo condiciones específicas del microambiente peritoneal, son capaces de implantarse y proliferar en forma ectópica. Se ha documentado que los macrófagos se encuentran aumentados en cuanto al porcentaje y grado de activación en ambiente peritoneal de mujeres con endometriosis. Se sabe que la activación de este grupo celular conduce a mayor síntesis de diversas moléculas que se asocian con el padecimiento. El objetivo de este estudio fue evaluar la asociación entre la capacidad de inducción de síntesis de óxido nítrico (NO) por parte del líquido peritoneal, el porcentaje de linfocitos T cooperadores y células NK presentes en ambiente peritoneal de mujeres con diversos estadios de endometriosis comparado con mujeres sanas fértiles. Además se buscó correlacionar la concentración de TNF-a identificado en el líquido peritoneal en ambos grupos de mujeres con la inducción de síntesis NO llevada. Material y métodos: se estudió grupo de mujeres con endometriosis (MEN) provenientes del Instituto Nacional de Perinatología, el grupo testigo fueron pacientes que acudieron a la Clínica de Planificación Familiar de la Unidad Regional del Noreste (Culiacán, Sin.) para que se realizara OTB (grupo MSF). Se realizó inducción de síntesis de NO en linfocitos estimulados con líquido peritoneal de MEN y MSF para analizar los linfocitos T cooperadores y NK, además, se determinó el TNF-a del líquido peritoneal. Resultados: se observó la capacidad del líquido peritoneal para inducir la síntesis de óxido nítrico por parte de linfocitos en cultivo es mayor con respecto a la observada en mujeres sanas. Conclusiones: se ha descrito recientemente que NO es un poderoso inhibidor de actividad efectora citotóxica asociada a respuesta inmunológica sobre linfocitos T cooperadores de tipo TH-1 que promueven actividad citotóxica sobre diversas estirpes celulares, al parecer el NO inhibe la síntesis de INF-a que es un potente inductor de proliferación y actividad citotóxica de células NK, linfocitos T citotóxicos y T cooperadores