Evaluation of agreement between conventional and liquid-based cytology in cervical cancer early detection based on analysis of 2,091 smears: experience at the Brazilian National Cancer Institute

The aim of this work was to evaluate the agreement between conventional cytology (CC) and liquid-based cytology (LBC) in cervical cancer early detection.The results of CC and LBC (DNACitoliq in 2,059 women aged 25-59 years were compared. The percent agreement, kappa coefficient, prevalence-adjusted bias-adjusted kappa coefficient (PABAK), and Chamberlain's percent positive agreement (PPA) were calculated.The percent agreement between the two methods was very good (80% and 79%, respectively, for normal versus ASCUS+; and normal versus ASCUS, AGUS and LSIL+ vs. HSIL+). The kappa coefficient indicates slight agreement (0.26 and 0.23, respectively), but when PABAK was used the agreement was good (0.61 and 0.68, respectively). PPA was high for normal results (79.2%) and low for the remaining categories.To conclude, in this study, agreement between LBC and CC was only good for normal results, which involves the majority of cases and positively influences the overall agreement rate.     

Brazilian Women With Lung Cancer Have a Higher Overall Survival Than Their Male Equivalents: A Cohort Study

BackgroundLung cancer is the principal cause of cancer-related deaths worldwide; however, there has been controversy as to whether there is a difference in survival rate according to gender in Brazil. The aim of the present study, therefore, was to compare the epidemiologic and clinical profile and the overall survival of patients with lung cancer according to gender.Patients and MethodsA retrospective cohort study was performed involving 1283 patients diagnosed with lung cancer between 2006 and 2014 at a single cancer center. Survival analysis was conducted using Kaplan-Meier statistics. A log-rank test was used to assess differences between survival curves, and Cox proportional hazards regression analysis was performed to quantitate the relationship between gender and overall survival.ResultsCompared with men, women were more frequently younger (P < .001), nonsmokers (P = .007), diagnosed with adenocarcinoma (P < .001), had early stage disease (P < .001), received surgery or surgery in combination with chemotherapy (P < .001), and had a better survival rate (P < .001). The median overall survival rate was higher in women (14.2 vs. 10.5 months in men; P < .001). Cox regression-adjusted analysis shows that women were 16% less likely to die than men (hazard ratio, 0.84; 95% confidence interval, 0.72-0.98; P = .03).ConclusionsA higher overall survival rate was found in women with lung cancer as compared with men with lung cancer in Brazil.     

Growth factors and stromal support generate very efficient retroviral transduction of peripheral blood CD34+ cells from Gaucher patients

We have achieved high-efficiency gene transfer into nonmobilized peripheral blood (PB) CD34+ cells from patients with Gaucher's disease using a clinically acceptable retroviral supernatant transduction protocol. In our studies, bone marrow (BM) and PB CD34+ cells were transduced using a high titer (10(8) particles/mL) retroviral supernatant once a day for 4 consecutive days in the presence of interleukin-3 (IL-3), IL-6, and stem cell factor (SCF), with or without an irradiated allogeneic BM stromal layer. The growth factors alone resulted in 29% +/- 10% gene transfer of PB CD34+ clonogenic cells in contrast with 71% +/- 17% gene transfer efficiency using stroma with the growth factors; a 2.5-fold increase. The increase in gene transfer efficiency was less prominent when BM CD34+ cells were used (40% +/- 16% without and 57% +/- 8% with stroma, a 1.5-fold increase). The overall transduction efficiency of both PB and BM CD34+ cells was lower when the cells were transduced over a stromal cell layer without added growth factors. The combination of IL-3, IL-6, and SCF with stroma transduced 75% of primitive long-term culture initiating cells (PB LTC- ICs) in comparison with 34% of LTC-ICs when IL-3, IL-6, and SCF were used without stromal support. Using this clinically acceptable supernatant/cytokines/stroma transduction protocol, correction of the glucocerebrosidase (GC) deficiency in the progeny cells of PBLTC-ICs from Gaucher's-disease patients has been accomplished. Efficient transduction of the PB CD34+ cells using this transduction protocol may allow repeated delivery of "GC-corrected" hematopoietic stem and progenitor cells to Gaucher's-disease patients.     

Direct demonstration that autologous bone marrow transplantation for solid tumors can return a multiplicity of tumorigenic cells

Patients with solid tumors are increasingly being treated by autologous bone marrow transplantation (BMT). Although response rates appear to be increased, disease recurrence is the commonest cause of treatment failure. Whether relapse is entirely due to residual disease in the patient or arises also from infiltrating malignant cells contained in the autologous marrow transplant has not been resolved. If the latter explanation is correct, then purging would be required as part of the transplantation procedure. We used retrovirally mediated transfer of the neomycin-resistance gene to mark BM harvested from eight patients with neuroblastoma in clinical remission. The marked marrow cells were subsequently reinfused as part of an autologous BMT. At relapse, we sought the marker gene in malignant cell populations. Three patients have relapsed, and in each the marker gene was detected by phenotypic and genetic analyses of resurgent malignant cells at medullary and extramedullary sites. Analysis of neuroblast DNA for discrete marker gene integration sites suggested that at least 200 malignant cells, each capable of tumor formation, were introduced with the autologous marrow transplant and contributed to relapse. Thus, autologous BMTs administered to patients with this solid tumor may contain a multiplicity of malignant cells that subsequently contribute to relapse. The marker-gene technique we describe should permit evaluation of the mechanisms of relapse and the efficacy of purging in patients receiving autologous marrow transplantation for other solid tumors that infiltrate the marrow.     

Translation and cross-cultural adaptation of the Multiple Sclerosis Walking Scale (MSWS-12) into Brazilian Portuguese

Poor walking performance is predictive of heart disease and osteoporosis and increases the risk of death in the elderly. Gait and vision have been identified as the most valuable physical functions according to multiple sclerosis patients' perceptions. The objective of this study was to perform a translation and cross-cultural adaptation of the Multiple Sclerosis Walking Scale (MSWS-12) into Brazilian Portuguese. A study of cross-cultural adaptation was conducted in ten steps. Participation in the study included four translators, two back-translators, twelve medical experts, twelve patients, twelve healthy subjects, and a Portuguese language expert. Only the question "Did standing make it more difficult to do things?" posed difficulty in the translation process. Maximum time for completion was less than three minutes (171 seconds). Internal consistency analyses showed high reliability (Cronbach's alpha = 0.94). The content validation and internal consistency stages were completed satisfactorily.     

Plasma P-selectin is increased in thrombotic consumptive platelet disorders

P-selectin is a 140-kD protein found in the alpha-granules of platelets and the Weibel-Palade bodies of endothelial cells that on cell activation is expressed on the cell surface and also secreted into the plasma. The secreted form of P-selectin, like plasma P-selectin, differed from platelet membrane P-selectin in that its molecular mass was approximately 3 kD lower under reducing conditions. Both the secreted and plasma forms of P-selectin contained cytoplasmic sequence as determined by Western blot analysis with an affinity-purified rabbit anti-P-selectin cytoplasmic peptide antibody. We have measured plasma P- selectin and beta-thromboglobulin (beta TG) concurrently in (1) patients with consumptive thrombotic disorders, including disseminated intravascular coagulation (DIC), heparin-induced thrombocytopenia (HIT), and thrombotic thrombocytopenic purpura (TTP)/haemolytic uremic syndrome (HUS); (2) patients with idiopathic thrombocytopenic purpura (ITP); and (3) healthy controls. Patients with DIC, HIT, and TTP/HUS, but not ITP, had significantly elevated plasma P-selectin and beta TG levels when compared with their age-matched healthy controls. The increased plasma P-selectin and beta TG in patients with thrombotic disorders were likely to be the result of in vivo platelet and endothelial cell damage or activation. We also found that avoidance of veno-occlusion and other tedious measures customarily taken during blood collection and sample preparation to prevent in vitro platelet activation did not affect plasma P-selectin assay results. In addition, plasma P-selectin levels were not influenced by the presence of renal failure or heparin administration. These results indicate that plasma P- selectin may be a useful new marker for thrombotic diseases.     

Three unrelated Rh D gene polymorphisms identified among blood donors with Rhesus CCee (r'r') phenotypes

Human red blood cells are traditionally typed as Rhesus (Rh)-positive or -negative depending on the presence or absence of the Rh D antigen. A recent report demonstrated that the Rh D gene is completely absent in Rh D-negative individuals. In this study, Rh D-negative blood donors with ccee (n = 25) and CCee (n = 3) phenotypes were examined for the presence of absence of the D gene. Polymerase chain reaction (PCR) probes that hybridize to the 5' and 3' regions of the Rh CcEe gene and the closely related D gene were used in a Southern analysis. The D gene was absent in all ccee phenotypes examined. The CCee phenotypes showed three Rh D polymorphisms: one donor lacked the D gene, one donor had a partial deletion on one D gene at the 3' region, and the remaining donor appeared to have one normal D gene within the intron/exon regions examined. We conclude that, while the D gene may be absent in the majority of Rh D-negative phenotypes, rarer polymorphisms also occur that prevent expression of the D antigen resulting in the Rh D-negative phenotype.     

B lymphoblastoid cell lines with normal and defective O-glycosylation established from an individual with blood group Tn

Individuals with the Tn blood group contain terminal serine/threonine- linked N-acetylgalactosamine residues in their blood cells. This is due to lack of UDP-D-galactose: D-N-acetyl galactosamine beta-D-galactosyl transferase from part of their red cells and probably from their leukocytes. We have established B lymphoblastoid cell lines from such an individual by in vitro infection of his lymphocytes with Epstein- Barr virus. The original line contained a mixture of cells reactive and nonreactive with Helix pomatia lectin (Hp). These cells were subcloned after staining with fluorescent Hp by a fluorescence-activated cell sorter (FACS) into homogeneous, phenotypically stable lines of Hp- positive (Hp+) and Hp-negative (Hp-) cells. The molecular differences between the membrane glycoproteins were characterized by carbohydrate- specific surface labeling techniques, Hp affinity chromatography, polyacrylamide slab gel electrophoresis and glycopeptide/oligosaccharide analysis. The major O-glycosidic membrane glycoprotein (GP105) was retained on Hp-Sepharose columns only from Hp+ cells, whereas the common leukocyte antigen (GP160-200) was partially retained on Hp columns from both lines. These proteins were isolated by immune precipitation with monoclonal antibodies and characterized. The results show that the GP105 glycoprotein from Hp+ cells contains terminal N-acetylgalactosamine residues but also more complex oligosaccharides. The common leukocyte antigen showed different electrophoretic mobilities in Hp+ and Hp- cells. UDP-galactose D-N- acetyl galactosamine beta-galactosyl transferase was almost absent in the Hp+ cells. These cell lines are useful for studies on the functional role and regulation of the biosynthesis of O-glycosidic carbohydrates.