Postoperative Depression of Tumour-directed Cell-mediated Immunity in Patients with Malignant Disease

Leucocytes from 46 melanoma patients, 45 breast carcinoma patients, and 95 control donors were tested by the leucocyte migration test against the supernatants of homogenates of malignant melanomas, breast carcinomas, simple breast tumours, and breasts showing simple cystic disease. By comparison with controls inhibition of migration occurred significantly more frequently when tumour patients'' leucocytes were exposed to extracts of histogenetically similar tumours.Cell-mediated immunity to tumour-associated antigens was measured in 12 patients with breast carcinoma and 12 with malignant melanoma immediately before surgical operation and in the postoperative period. All patients tested before operation showed significant inhibition of migration on contact with extracts of histogenetically similar tumours. Postoperatively the degree of leucocyte migration inhibition was reduced in all patients with melanoma and breast carcinoma. Significant inhibition of leucocyte migration returned in most patients 6-22 days after operation.     

Primary Hepatoma and Hepatitis-associated Antigen in a Young White Woman

A young woman whose serum was positive for hepatitis-associated antigen (H.A.A.) and alpha-fetoprotein developed a malignant hepatoma. Though the light-microscopical appearances of the surrounding liver tissue were normal, electronmicroscope examination of the tumour tissue disclosed both C-virus-like particles and H.A.A. particles. Possibly there was a causal connexion between one of these particles and the hepatoma.     

Immunological aspects of benign melanocytic tumours and melanoma in situ

Summary Extracts were prepared from 15 malignant melanomas, 10 naevi, 8 pieces of skin containing atypical melanocytes and 4 samples of normal skin. Peripheral blood leucocytes from 52 patients with benign naevi that had altered sufficiently to cause the patient to seek attention, 57 patients with malignant melanomas, and 98 normal individuals were tested against these extracts in a capillary leucocyte migration test. Melanoma extracts preferentially inhibited the leucocytes of melanoma patients and to a lesser extent those of naevus patients. Melanoma patients' leucocytes were more frequently inhibited by naevus extracts (whether from intradermal or from compound naevi) than were the leucocytes of naevus patients and control donors. Extracts of skin containing atypical melanocytes (perimelanomatous skin and lentigenous skin) selectively inhibited melanoma patients' leucocytes.     

Evaluation of the Gonosticon Dri Dot Test in Females with a Low Incidence of Gonorrhea

A group of 765 females attending a Planned Parenthood Clinic was screened for gonorrhea by inoculating Thayer-Martin plates and Transgrow bottles with specimens from the cervix. Blood was obtained at the same time and tested for anti-gonococcal antibody by using the Gonosticon Dri Dot test. In this low-incidence group, 18 positive cultures were detected by culture on Thayer-Martin plates, whereas Transgrow detected only 15 positive cultures. Of the 18 patients with gonorrhea, 11 exhibited reactive serum (agglutination of the latex particles). In the total population, 64% of the patients had nonreactive serum (no agglutination) and negative cultures; 25% had reactive serum and negative cultures. When this latter group was subdivided on the basis of race, blacks and Latin Americans were found to have a higher incidence of reactive serum with a corresponding negative culture than was found in whites. Patients who were originally culture positive and nonreactive in the Gonosticon Test were retested; three out of four patients retested within 6 to 11 days after the initial screening had converted to a positive Gonosticon test.     

Detection of HCV-RNA in liver tissue does not correlate with inflammatory process and fibrosis activity in chronic hepatitis C

The significance of HCV-RNA presence in the liver tissue in chronic hepatitis C activity or prognosis has not yet been clearly explained. Therefore, we have examined the relationship between the presence of HCV-RNA in the liver and selected parameters of disease activity and liver damage. A group of 48 chronically HCV infected patients (7-63 years old, mean 39 years) was evaluated. In the patients we assessed the activity of transaminases (ALT, AST), gammaglutamyltransferase (GGTP), and alkaline phosphatase (ALP). The patients underwent routine liver biopsies and the liver tissue was examined histopathologically and in order to detect the presence of HCV-RNA, by means of a combined procedure joining a new method of HCV-RNA extraction from the liver tissue and HCV-RNA detection with RT-PCR automatic Cobas Amplicor Hepatitis C ver. 2.0 assay (Roche Diagnostics). At the time of the liver biopsy, 44 of the patients were identified as having detectable serum HCV-RNA (as examined by means of Cobas Amplicor Hepatitis C ver. 2.0 assay), 3 patients were negative, and 1 was not tested. The presence of HCV-RNA in the liver tissues was detected in 39 cases (81.2%). In the parameters examined we have not found any significant differences between currently liver HCV-RNA positive and negative patients. Presence of the detectable HCV-RNA in the liver bioptats from chronic hepatitis C patients does not correlate with disease activity or level of liver damage.     

Detection of benign or malignant origin of ascites with combined indirect immunoperoxidase assays of carcinoembryonic antigen and lysozyme

The indirect immunoperoxidase method was used to study the presence of the intracellular carcinoembryonic antigen (CEA) and lysozyme (LZ) in alcohol-fixed cytologic smears of peritoneal fluids from 2 patients with chronic active hepatitis, 31 patients with liver cirrhosis and 7 patients with malignant liver disease. In the two patients with hepatitis, LZ was positive in both CEA was positive in one and negative in the other. Of the 31 patients with liver cirrhosis, 21 (67.5%) were LZ positive, 27 (87%) were CEA negative and only 4 (13%) were CEA positive. Of the seven patients with malignant disease, six were CEA positive and six were LZ negative. It is of interest that 23 of 24 (96%) LZ-positive results and 28 of 29 (97%) CEA-negative results corresponded to negative cytologic diagnoses for malignancy. Cytologic diagnosis of "reactive mesothelial cells" seemed to correlate better (71%) with CEA-negative and LZ-positive results. The data suggest that the investigation of CEA and LZ in the cells of peritoneal fluids appears to have promise as an adjunct to cytology in differentiating benign from malignant origins of the fluid.     

Hepatitis B virus (HBV) DNA in leucocytes in acquired immune deficiency syndrome (AIDS)

Earlier reported findings of hepatitis B virus (HBV) DNA in white blood cells of patients with hepatoma, and in a patient with autoimmune haemolytic anaemia, led to the examination of HBV DNA in a series of twenty three patients with acquired immune deficiency syndrome (AIDS), including nine with opportunistic infections and fourteen with Kaposi's sarcoma, by Southern blot hybridization method, using 32P labelled HBV DNA specific probe obtained by nick translation of HBV DNA cloned into plasmid pBR325. Four of the patients were found to be positive for HBV DNA or HBV related DNA in their leucocytes. The HBV DNA was found free or integrated in the leucocytes of the patients.     

泛素相关蛋白样因子2在肝癌中的表达及意义

目的:探讨肝癌中泛素相关蛋白样因子2(UBAP2L)的表达水平与预后的关系及其对肝癌细胞侵袭、转移能力的影响。方法:挖掘oncomine数据库,提取UBAP2L在肝癌与正常组织转录水平的变化及相关临床资料,采用Kaplan-Meier法分析UBAP2L表达水平与肝癌患者预后的关系。采用实时定量PCR、Western blot检测HCCLM3、MHCC97H、Hep3B、Huh7肝癌细胞株及正常肝细胞LO2中UBAP2L mRNA及蛋白水平,免疫组织化学染色法检测本院80例肝癌组织与癌旁组织中UBAP2L的表达水平加以验证。通过慢病毒载体使UBAP2L高表达的肝癌细胞株HCCLM3表达下调,采用克隆形成和划痕实验检测UBAP2L对肝癌细胞增殖能力和侵袭、转移的影响。结果:UBAP2L在oncomine数据库大多数队列呈高表达。UBAP2L在肝癌细胞株中的mRNA和蛋白的表达水平显著高于正常肝细胞(P0.05)。肝癌组织中UBAP2L阳性、强阳性表达率高于癌旁组织(P0.05),下调UBAP2L表达可明显抑制HCCLM3肝癌细胞的克隆形成能力和运动能力(P0.05)。UBAP2L高表达组的中位生存时间与UBAP2L低表达组比较差异无统计学意义(P0.05)。结论:UBAP2L在肝癌组织中呈高表达,下调UBAP2L表达可抑制肿瘤增殖和侵袭、转移表型,其可能成为肝癌早期诊断的生物标志物和治疗的潜在靶点。     

Relevance of Non-ceruloplasmin Copper to Oxidative Stress in Patients with Hepatocellular Carcinoma

Altered copper homeostasis and oxidative stress have been observed in patients with hepatocellular carcinoma. Non-ceruloplasmin copper, the free form, is a potent pro-oxidant than the protein bound copper. The aim of the present study was to evaluate which form of copper can be correlated with the oxidative stress in the circulation and in the malignant liver tissues of hepatocellular carcinoma patients. Hepatocellular carcinoma patients (grades II and III, n = 18) were enrolled in this study. Serum levels of total, free and bound copper, ceruloplasmin, iron, iron-binding capacity, lipid peroxidation products, and enzymatic and non-enzymatic antioxidants were quantified in serum and in malignant liver tissues and compared with those of normal samples (n = 20). A significant positive correlation between the serum non-ceruloplasmin copper and lipid peroxidation products and negative correlation with antioxidants were observed in hepatocellular carcinoma patients. In liver tissue, glutathione peroxidase, superoxide dismutase, and catalase activity were significantly decreased with concomitant elevation in oxidative stress markers. Our experiment revealed that the elevation in non-ceruloplasmin copper has high relevance with the oxidative stress than the bound copper.     

Features of the biosynthesis of immunoglobulin G peculiar to the growth process

It is established that with partial hepatectomy, Shvets leukosis and hepatoma RS-1 the biosynthesis intensity of rat blood serum proteins producing aggregates in the acid medium is considerably higher than that of other serum proteins, the incorporation of the radioactive precursor into immunoglobulin G peculiar to intensive normal and malignant growth being particularly intensive. During liver regeneration as well as in malignant growth specific radioactivity of immunoglobulin G peculiar to the growth processes is three and five times, respectively, as high as this value for blood serum soluble proteins and proteins of alpha-globulin fractions.     

Serum screening for Down's syndrome: some women's experiences.

OBJECTIVES--To describe the experiences of a small group of women who had positive results after serum screening for Down''s syndrome. DESIGN--Semistructured telephone interviews and correspondence with women after a positive screening result (four women) negative amniocentesis results (eight), or termination of a pregnancy with a confirmed abnormality (eight). SUBJECTS--20 women who contacted Support After Termination For Abnormality about their experiences of serum screening for Down''s syndrome. MAIN OUTCOME MEASURES--Women''s knowledge and understanding of the test; staff misconceptions; communication of results; how women coped with the diagnostic process; attitudes to the test and to termination of abnormal fetuses. RESULTS--All women were made anxious by their positive screening test, no matter how they were told. The women''s experiences suggested that medical staff were unclear about the implications of screening tests and how to interpret risk. Even after receipt of negative amniocentesis results some women remained anxious. Staff did not always recognise women''s concerns while awaiting amniocentesis results. CONCLUSIONS--The way in which serum screening is being implemented does not always meet the needs of women with positive results. Some of the problems were not specific to screening for Down''s syndrome. When screening tests are introduced policies should be adopted to ensure appropriate support for participants.     

Immunocytochemical localization of testosterone in human hepatocellular carcinoma

Summary A study of 10 hepatoma patients was carried out for the detection and localization of oestrogen, progesterone, testosterone, and their corresponding binding sites in liver tumour cells using immunoperoxidase and immunofluorescent techniques on histopathological sections and cultured monolayers. None of the hepatomas was positive for progesterone and its receptor; one was positive for oestrogen and its receptor; five were positive for testosterone and its receptor. Oestrogen, testosterone and their respective receptors localized predominantly to the cytoplasm of hepatoma cells. The results obtained in this investigation reveal an association between testosterone and human liver cancer which may be useful in the selection and monitoring of liver cancer patients to future hormonal therapy.     

Immunodiagnosis of ocular toxocariasis using Western-blot for the detection of specific anti-Toxocara IgG and CAP for the measurement of specific anti-Toxocara IgE

A prospective multicentric study was carried out to assess both the performance of Western-blot (WB) detecting specific anti-Toxocara IgG and that of CAP measuring specific IgE titre for the immunodiagnosis of ocular toxocariasis. For 14 outpatients presenting ophthalmic symptoms (choroiditis, chorioretinitis, papillar oedema, hyalitis, retinal detachment and/or uveitis), samples of serum and aqueous fluid (AF) were sent to the Department of Parasitology, University Hospitals, Toulouse, France. All patients but two tested positive with WB on the serum; 13 WB tests were performed on the AF, 12 of which were positive. The two patients who had a negative WB serum result tested positive for the AF. Specific IgE detection was considered as a complementary test of WB. Two patients showed a greater specific IgE titre in the AF than in the serum, and one had a positive result in the AF, but not in the serum. These six patients were considered as clear cases of ocular toxocariasis. Western-blot coupled with specific anti-Toxocara IgE detection appeared therefore to be an accurate procedure for the immunodiagnosis of ocular toxocariasis, provided the testing was simultaneously performed on the serum and AF.     

Evaluation of an enzyme immunoassay for hepatitis C virus antibody detection using a recombinant protein derived from the core region of hepatitis C virus genome

This study was undertaken to evaluate an enzyme immunoassay (EIA) for hepatitis C virus antibody detection (anti-HCV), using just one antigen. Anti-HCV EIA was designed to detect anti-HCV IgG using on the solid-phase a recombinant C22 antigen localized at the N-terminal end of the core region of HCV genome, produced by BioMérieux. The serum samples diluted in phosphate buffer saline were added to wells coated with the C22, and incubated. After washings, the wells were loaded with conjugated anti-IgG, and read in a microtiter plate reader (492 nm). Serum samples of 145 patients were divided in two groups: a control group of 39 patients with non-C hepatitis (10 acute hepatitis A, 10 acute hepatitis B, 9 chronic hepatitis B, and 10 autoimmune hepatitis) and a study group consisting of 106 patients with chronic HCV hepatitis. In the study group all patients had anti-HCV detected by a commercially available EIA (Abbott), specific for HCV structural and nonstructural polypeptides, alanine aminotransferase elevation or positive serum HCV-RNA detected by nested-PCR. They also had a liver biopsy compatible with chronic hepatitis. The test was positive in 101 of the 106 (95%) sera from patients in the study group and negative in 38 of the 39 (97%) sera from those in the control group, showing an accuracy of 96%. According to these results, our EIA could be used to detect anti-HCV in the serum of patients infected with hepatitis C virus.     

Chlamydia trachomatis infection in rural Nova Scotia.

OBJECTIVE: To examine the demographic characteristics of patients who underwent testing for Chlamydia trachomatis and to determine the clinical and behavioural characteristics and the types of treatment for those who had positive test results. DESIGN: Case series. SETTING: Rural county in Nova Scotia. PATIENTS: All residents of the county for whom testing for C. trachomatis was ordered at the regional hospital from Sept. 1, 1990, to Mar. 31, 1991. MAIN OUTCOME MEASURES: Rates of testing and of positive test results by age and sex. Comparison of patient and physician characteristics in relation to testing rates. RESULTS: Of the 1116 patients tested 58 (5.2%) had positive test results. Females accounted for 82.8% of those with positive results whose sex could be determined. Among the females the mean age of those with a positive result was 22.3 years, as compared with 27.5 years for those with a negative result (p < 0.0001). Females 15 to 19 years of age were less likely to have a test performed than women 20 to 29 years and were more likely to have a positive test result than the women in the older groups. Almost 9% of the testing among the females was in those over 39 years of age, although no infection was seen in this age group. The number of tests ordered per general or family practitioner varied from 1 to 154; the physicians'' sex, practice location and length of time in practice did not predict the rates of positive test results. Treatment was most often in keeping with that recommended by national guidelines. Four (8.5%) of the 47 patients with positive results who were interviewed were not aware of their diagnosis, either because they had not returned for follow-up or had not being notified by the physician''s office. CONCLUSIONS: The frequency of testing for C. trachomatis infection may be less than is desirable among young patients, who, if tested, are more likely than older patients to have positive results. More understanding of the diagnostic approach taken by physicians is needed.     

A study of the cell surface of tumour, foetal and lymph-node cells by cell electrophoresis after antibody and enzymic treatment

1. Rabbit anti-(rat foetal liver) serum, absorbed with adult rat liver cells, decreased the electrophoretic mobility of foetal liver cells by 51% and rat hepatoma cells by 45%, indicating the presence of a foetal-type antigen on the hepatoma cell membrane. 2. The chemical nature of the surface antigen was investigated. Incubation with neuraminidase had no effect on adult liver cells but decreased the electrophoretic mobility of foetal liver cells by 51% and of hepatoma cells by 34%; the effect of antiserum was decreased to one-fifth. 3. Sialic acid, or the supernatant from neuraminidase-treated cells, partially blocked the decrease in electrophoretic mobility induced by antiserum. 4. The pH-electrophoretic mobility curves of hepatoma cells treated with antisera were consistent with a sialic acidcontaining antigen on the surface of the tumour cells. 5. Treatment with ribonuclease did not decrease the electrophoretic mobility of adult-liver cells, but decreased that of the foetal liver cells by 17% and hepatoma cells by 29%. 6. In parallel studies made with mouse BP8 ascites-tumour cells ribonuclease decreased the electrophoretic mobility by 39%, that of normal mouse lymph-node cells by 4.8% and allergized mouse lymph-node cells by 13.3%. 7. Trypsin treatment also decreased the electrophoretic mobility of hepatoma cells by 22%.     

Impact of the Use of a Rapid Diagnostic Test for Visceral Leishmaniasis on Clinical Practice in Ethiopia: A Retrospective Study

Background

Diagnostic guidelines for Visceral Leishmaniasis (VL) in the East African region are complex. Patients meeting the VL clinical case definition should be tested by rK39 rapid diagnostic test (RDT) followed by the Direct Agglutination Test (DAT) or tissue aspiration if RDT-negative. Otherwise, RDT-positive patients should be started on VL treatment. We evaluated how this guideline is adhered to by assessing the routine clinical practice in a university hospital in North-West Ethiopia.

Methods

Retrospective record analysis was done for all patients who had an rK39-RDT done at University of Gondar (UoG) Hospital between June 2012 and June 2013. We described the diagnostic work-up performed and the proportion initiated on VL treatment by test result.

Results/Findings

From a total of 928 patients tested, 308 (33.2%) were rK39 RDT-positive. Spleen or bone marrow aspiration was done for 237 (77.2%) RDT-positive patients. Of these, 165 were confirmed parasitologically, yielding a positive predictive value of 69.6%. Only 126 (20.3%) of the 620 patients with a negative rK39 test underwent further testing by tissue aspiration, of which 22 (17.5%) were also parasitology positive. HIV test results were available for 570 (61.4%) patients and 36 (6.3%) were HIV-infected. Of the 187 parasitologically confirmed patients, 182 (97.3%) were started on VL treatment.

Conclusions / Discussion

A negative rK39 test was often not followed by further testing and a positive rK39 test result was followed by tissue aspiration in three out of four cases. Further research is required to understand why the diagnostic work-up did not comply with the guidelines, including evaluating adherence to the VL clinical case definition and quality of rK39-RDT testing.     

Studies on DNA content of hepatocytes in cirrhosis and hepatoma by means of microspectrophotometry and radioautography

In order to clarify the underlying mechanism of malignant transformation from cirrhosis to hepatoma the cell kinetics of hepatocytes were studied in these two conditions. The content and synthesis of DNA in hepatocyte nuclei were investigated, by means of Feulgen-microspectrophotometry and tritiated thymidine radioautography, in cirrhotic and noncancerous parts of hepatoma with concomitant cirrhosis. The distribution of ploidy patterns was widely spread, from hypodiploid to hyperpolyploid, in the noncancerous parts of a cirrhotic liver containing hepatoma. In normal liver, each paired nuclear DNA content of a binucleate cell recorded almost the same amount, whereas in the noncancerous as well as in hepatoma cells much difference of DNA content was observed between the paired nuclei of the binucleate cells. The ploidy pattern of hepatocytes in patients with liver cirrhosis, who had developed hepatoma during follow-up periods of several months to several years, appeared to resemble that in noncancerous parts of hepatoma cases. On the other hand, the incorporation of tritiated thymidine into hepatocytes was found to be markedly increased in noncancerous parts as well as in cirrhotic liver developing hepatoma during follow-up periods. These results suggest the possibility that the hepatocytes in noncancerous parts of hepatoma have deranged cell-kinetics which might be a driving factor for the development of malignancy.     

Studies on DNA content of hepatocytes in cirrhosis and hepatoma by means of microspectrophotometry and radioautography

Summary In order to clarify the underlying mechanism of malignant transformation from cirrhosis to hepatoma the cell kinetics of hepatocytes were studied in these two conditions. The content and synthesis of DNA in hepatocyte nuclei were investigated, by means of Feulgen-microspectrophotometry and tritiated thymidine radioautography, in cirrhotic and noncancerous parts of hepatoma with concomitant cirrhosis. The distribution of ploidy patterns was widely spread, from hypodiploid to hyperpolyploid, in the noncancerous parts of a cirrhotic liver containing hepatoma. In normal liver, each paired nuclear DNA content of a binucleate cell recorded almost the same amount, whereas in the noncancerous as well as in hepatoma cells much difference of DNA content was observed between the paired nuclei of the binucleate cells. The ploidy pattern of hepatocytes in patients with liver cirrhosis, who had developed hepatoma during follow-up periods of several months to several years, appeared to resemble that in noncancerous parts of hepatoma cases. On the other hand, the incorporation of tritiated thymidine into hepatocytes was found to be markedly increased in noncancerous parts as well as in cirrhotic liver developing hepatoma during follow-up periods. These results suggest the possibility that the hepatocytes in noncancerous parts of hepatoma have deranged cell-kinetics which might be a driving factor for the development of malignancy.     

Levels of hepatitis B virus replicative intermediate in serum samples of chronic hepatitis B patients

Hepatitis B virus (HBV) cccDNA levels is an absolute marker of HBV replication in the liver of HBV infected patients. This study aimed to quantify the HBV cccDNA levels in sera and liver tissue samples of treatment naïve patients with chronic hepatitis B. Eighty one chronic hepatitis B (CHB) treatment naïve patients were enrolled from January 2009 to June 2011. Total HBV DNA and HBV cccDNA levels were quantified using sensitive real time PCR assay. The mean age of recruited patients was 34 ± 11.5 years. Fifty four (66.7 %) patients were HBeAg negative. Liver tissue samples were available from 2 HBeAg positive and 21 HBeAg negative CHB patients. The amount of total intrahepatic HBV DNA ranged from 0.09 to 1508.92 copies/cell. The median intrahepatic HBV cccDNA was 0.31 and 0.20 copies/cell in HBeAg positive and HBeAg negative cases, respectively. Serum HBV cccDNA was detectable in 85.2 % HBeAg positive and 48.1 % HBeAg negative CHB patients. Median serum HBV cccDNA was 46,000 and 26,350 copies/mL in HBeAg positive and HBeAg negative subjects, respectively. There was a significant positive correlation between the levels of intrahepatic total HBV DNA and intrahepatic HBV cccDNA (r = 0.533, p = 0.009). A positive correlation was also seen between serum HBV cccDNA levels and serum HBV DNA levels (r = 0.871, p < 0.001). It was concluded that serum HBV cccDNA could be detectable in higher proportion of HBeAg positive patients compared to HBeAg negative patients. Moreover, the median level of serum HBV cccDNA was significantly higher in HBeAg positive patients in contrast to HBeAg negative subjects.