Kirsten murine sarcoma virus abolishes interferon gamma-induced class II but not class I major histocompatibility antigen expression in a murine fibroblast line.

The effect of infecting fibroblasts with Kirsten murine sarcoma virus/murine leukemia virus (Ki-MSV/MLV) on constitutive and IFN-gamma-induced H-2 antigen expression was investigated. The fibroblasts used were two established cell lines (C3H10T1/2 and BALB/c3T3) and fresh embryo fibroblasts from C3H mice. Class I antigens were expressed constitutively by BALB/c3T3; infection with MLV, MSV or the two together had little effect on this constitutive expression. Class I antigens (H-2K, H-2D) were strongly induced on all three types of fibroblast by rIFN-gamma, and infection had little effect on this. None of the fibroblasts expressed constitutively detectable levels of class II antigen; however, C3H10T1/2 fibroblasts could be induced for both H-2A and H-2E by IFN-gamma. Infection of C3H10T1/2 with helper-free Ki-MSV, or MSV together with MLV, completely abolished this induction of class II antigens, while infection with MLV alone had little effect, implying that the abolition of class II induction was due to genomic regions of Ki-MSV not shared with Ki-MLV, probably the v-Ki-ras gene.

Inducibility of class II major histocompatibility complex antigens by interferon gamma is associated with reduced tumorigenicity in C3H mouse fibroblasts transformed by v-Ki-ras.

Paired lines of C3H mouse fibroblasts transformed with murine sarcoma virus (Kirsten strain) were prepared that express high or low levels of class II major histocompatibility complex antigen after treatment with interferon gamma (IFN-gamma). Here, we described a comparison of the tumorigenicity of these lines in euthymic syngeneic and thymus-deficient nu/nu mice and in mice depleted of IFN-gamma. The class II-inducible cells are clearly less tumorigenic than the noninducible cells in syngeneic mice, but of similar tumorigenicity in nu/nu mice and in mice treated with antibodies to deplete IFN-gamma. We propose that in this system, IFN-gamma induction of class II antigens on the tumor cell surface operates to limit tumor growth; ras expression, which inhibits induction of class II antigens, prevents this and so allows tumor growth.

Effect of dopamine D3 receptor gene polymorphisms and clozapine treatment response: exploratory analysis of nine polymorphisms and meta-analysis of the Ser9Gly variant.

D2 blockade has been implicated in having a central role in antipsychotic response. However, treatment refractoriness, in spite of complete D2 blockade, as well as the efficacy of clozapine (CLZ) in a portion of this patient population, indicates the involvement of other factors as well. Several lines of evidence suggest a role for D3. Furthermore, an earlier meta-analysis by Jönsson et al. (2003) (n=233) suggested a role for genetic variation in the D3 gene. Relevant to this study, Jönsson et al. found the Ser allele of the D3 serine-to-glycine substitution at amino acid position 9 (Ser9Gly) polymorphism to be associated with worse CLZ response compared with the Gly allele. In this study, we attempt to validate these findings by performing a meta-analysis in a much larger sample (n=758). Eight other variants were also tested in our own sample to explore the possible effect of other regions of the gene. We report a negative but consistent trend across individual studies in our meta-analysis for the DRD3 Ser allele and poor CLZ response. A possible minor role for this single-nucleotide polymorphism cannot be disregarded, as our sample size may have been insufficient. Other DRD3 variants and haplotypes of possible interest were also identified for replication in future studies.

Precocious natural mummification in a temperate climate (Western Cape, South Africa).

The general process and pattern of decomposition is well-documented and understood. However, specific environmental conditions may alter this pattern and prematurely terminate the decay process. An example of this is natural mummification - a preservative process characterized by desiccation, brittleness and shrinkage of the skin and body tissues. It is important to understand how, when, and where such variations may occur, and for this reason environmentally-specific studies of decay are required. The aim of the present study was the establish baseline data on soft-tissue decomposition in two terrestrial habitats in the Western Cape. A total of 16 pig carcasses serving as analogues for humans were deployed in these habitats during two successive winters and summers between 2014 and 2016. The rate and pattern of decomposition were assessed via measurement of weight loss over time and scoring the decomposition process using Megyesi et al. (2005) Total Body Score system and study-specific criteria for mummification. Carcasses typically followed the expected pattern of decay with a few exceptions, most notably instances of rapid natural mummification. Natural mummification, as defined by Megyesi et al. (2005), was observed to occur as early as 17 days postmortem, with five carcasses mummifying in less than one month. The timing of natural mummification varies widely, from a few days to several years, averaging around three months in temperate regions. Natural mummification occurring in less than one month is termed precocious mummification and is rarely observed in temperate regions. With only three reports globally, this study's findings are globally significant, highlighting the importance of regionally-specific decomposition studies. Two local forensic cases wherein precocious mummification has been observed are also presented and, considered together with the study's results, a possible mechanism driving this process is proposed.

The prognostic and predictive value of syntactic structure analysis in serous carcinoma of the ovary.

The objective of this study was to determine whether syntactic structure analysis (SSA) can predict survival outcome and chemotherapeutic response in ovarian carcinoma. Syntactic structure analysis parameters, blindly determined in archived hematoxylin and eosin sections of 132, International Federation of Gynecology and Obstetrics (FIGO) stage I to IV serous ovarian tumors, and clinicopathologic parameters were evaluated as to their univariate and multivariate prognostic value and ability to predict chemotherapy response as measured by changes in CA125 levels. Univariate analysis revealed FIGO stage, tumor grade, preoperative CA125, presence of ascites, extent of disease residuum, and the SSA parameters minimum spanning tree (min MST), maximum MST (max MST), percent connectivity to 1, and 2 nearest neighbors to be significant predictors of overall survival and disease-free survival. Tumor grade, FIGO stage, extent of disease residuum, presence of ascites, and percent connectivity to 2 nearest neighbors were found to be significant predictors of chemotherapy response. Multivariate analysis revealed extent of disease residuum to be a significant predictor for overall survival (P

The prognostic value of nuclear morphometric analysis in serous ovarian carcinoma.

The objective of this study was to determine whether nuclear morphometric data can predict survival, disease progression, and chemotherapeutic response in ovarian serous carcinoma. Nuclear morphometric parameters were determined from archival hematoxylin and eosin sections of 132 serous tumors. Clinicopathologic and morphometric parameters were evaluated as to their individual and independent prognostic value and prediction of chemotherapy response. Nuclear parameters were found to strongly correlate with extent of disease residuum, tumor grade, and FIGO stage. Univariate analysis revealed stage, grade, preoperative CA125, presence of ascites, extent of disease residuum, standard deviation of nuclear area (SDNA), nuclear perimeter (NP), SDNP, nuclear length (NL), nuclear breadth (NB), orthoferet, and equivalent diameter (ED) to be significant predictors of overall survival (OS) and disease-free survival (DFS). Grade, stage, extent of disease residuum, presence of ascites, SDNA, NP, NL, NB, and orthoferet were found to be significant predictors of chemotherapy response. Multivariate analysis revealed extent of disease residuum (P

P53 and bcl-2 assessment in serous ovarian carcinoma.

The study objective was to determine the prognostic value of assessment of staining of p53 and bcl-2 in a well-selected group of serous ovarian carcinomas. Immunohistochemical detection was used to identify both p53 and bcl-2 positive tumors. One hundred thirty-two tumors were analyzed for positivity of staining, grade of staining intensity, and for p53 alone, percent expression rates. These were analyzed alongside traditional clinicopathologic parameters for their ability to predict overall survival (OS), disease-free survival (DFS), and response to chemotherapy (CR). Univariate COX analysis revealed percent p53 expression (P = 0.012) and p53 grade (P = 0.01) to be significant predictors of DFS. Neither the p53 nor bcl-2 measurement parameters were found significant for OS or prediction of CR. On multivariate analysis, incorporating clinicopathologic parameters, p53 parameters did not retain independent significance for any outcome measure. As in primary reported studies, bcl-2 was not found to be of clear independent prognostic value in this group of ovarian tumors. If mutation of p53 and its consequent overexpression is an early event in ovarian tumorigenesis, then p53 assessment may prove useful prognostically in the assessment of either low-grade ovarian carcinomas, as a possible indicator for progression, or in early-stage ovarian tumors, as a marker of tumor aggression or likelihood of recurrence. p53 analysis of a larger group of stage I ovarian tumors would be desirable to further explain the potential association with DFS.

Pulling teeth for fashion: dental modification in modern day Cape Town, South Africa.

Friedling and Morris (2005) have reported that intentional removal of incisors as a form of dental modification is relatively common in Cape Town. In this paper we further report on the style of modification and the reasons for the modification. A survey of eight adjoining areas in the northern suburbs of the Cape Town Metropole in the Western Cape was done to investigate the current prevalence of this practice. The survey was conducted by means of a questionnaire. Three groups of study subjects (scholars, working people and retired people) were included to gain a perspective of the community in general. The individual ages ranged from 15 to 83-years-old. A total of 2167 individuals participated in this study. Forty one percent had modified their teeth. More males (44,8%) than females (37,9%) were involved in this practice. Six "styles" of modification were identified. The removal of the upper four incisors was by far the most common modification (93,7%). There were four reported reasons for dental modification i.e. gangsterism, peer pressure, fashion and medical (dental) or accidental. More than two thirds (69,8%) of individuals with modifications also wore dentures.

Common variants in the chromosome 2p23 region containing the SLC30A3 (ZnT3) gene are associated with schizophrenia in female but not male individuals in a large collection of European samples.

Previously, we found a significant gender-specific association of schizophrenia, in a UK case/control study, with SLC30A3, a candidate that is consistently down-regulated in schizophrenia in two independent cohorts. In view of the potential significance of this finding, we extended this study to a larger cohort using GWAS data from the Psychiatric Genetic Consortium (PGC). Meta-analysis was performed for the only two SLC30A3 SNP variants (rs11126936 and rs11126929) available in most PGC cohorts. A significant association with schizophrenia was found for both variants. When meta-analysis was performed in male and female case-control subsets, an increased and gender-specific effect of allele on risk of disease was found in females for both SNPs with no significant effect in males, which was further associated with a gender-specific effect on gene expression. In conclusion, using a large European-wide sample we were able to replicate the gender-specific association previously found in a UK cohort.

The frequency of culturally derived dental modification practices on the Cape Flats in the Western Cape.

The people living on the Cape Flats (Western Cape) have been practicing dental modification by having their incisors removed for at least 60 years. A survey, by means of a questionnaire, of eight adjoining areas in the Northern Suburbs was done to investigate the current prevalence of this practice. Three groups of study subjects (scholars, working people and retired people) were included to gain a perspective of the community in general. The individual ages ranged between 15 and 83 years old. A total of 2 167 individuals participated in this study of which 41% had modified their teeth. More males (44.8%) than females (37.9%) were involved in this practice. The percentage incidence was overwhelmingly higher in lower income areas. 'Race' and social class are both important factors. Of people who called themselves 'coloured', 74.5% modified their teeth, but 39.8% of 'blacks' and 31.6% of 'whites' within the sample had also practiced tooth removal. The frequency of tooth removal for those who identified themselves as South African was much lower (22.9%). This suggests that dental modification is a phenomenon of ethnicity and community and those who perceive themselves to be part of a broader national fabric are less likely to practice it.

Educational intervention for preschool children in a pediatric clinic.

A parent education program aimed at promoting cognitive development in 2- and 3-year-olds was instituted in both a pediatric clinic of a general hospital and a child health station in the community. Parent participants were asked to work at home daily with their children using age-graded play materials supplied by the program. They were tutored individually, in the clinic playroom, by bilingual playroom assistants using role-playing techniques. One hundred fifty children completed the program out of a total of 468. Subjects were tested before and after training on the Cattell: Stanford-Binet and showed a positive gain in performance. Parents reported that their program experiences made them aware of their ability to teach their children. This was reflected in their changing view of the children's activities which were previously seen as aimless and "bad" and were now viewed as constructive and amenable to modification by the parents.

Accessory cell requirements for T lymphocyte activation and interferon-gamma production in peripheral lymph nodes.

Allergen-activated draining lymph node cells (LNC) isolated from mice exposed topically to the contact allergen oxazolone mount vigorous proliferative responses and secrete substantial amounts of interferon-gamma (IFN-gamma) when cultured with the T lymphocyte mitogen concanavalin A (con A). In contrast, although naive LNC prepared from untreated mice display con A-driven proliferative responses of comparable magnitude, they produce only very low levels of IFN-gamma. Secretion of IFN-gamma by con A-stimulated naive LNC was augmented significantly by the addition to culture of a small number of syngeneic dendritic cells (DC), under conditions where there was no influence on the vigour of proliferative responses and where the exogenous DC themselves failed to produce IFN-gamma. Augmentation of IFN-gamma production was not observed when exogenous populations depleted of DC were added to culture. It is proposed that discrete aspects of the primary activation of naive T lymphocytes display differential requirements for accessory cells and that the development of IFN-gamma producing cells necessitates sufficient numbers of dendritic cells.

The number of triplet repeats in five brain-expressed loci with CAG repeats is not associated with schizophrenia.

We have previously shown that large expansions of CAG (CTG) triplets are associated both with schizophrenia itself and with an early age-at-onset of the disease. However, the repeat expansion detection (RED) method used did not provide a chromosomal location for the expanded region(s) (Morris et al., 1995). In a further study of our schizophrenic and control patients, we have now examined the length of the repeated sequence in five loci that are expressed in brain and are known to contain CAG repeat regions (Li et al., 1993). No enlarged repeat regions were identified; it is unlikely therefore that expansions at any of these five loci can account for expansions of up to 136 triplets identified by the RED method.

Expression of HLA-ABC, HLA-DR and intercellular adhesion molecule-1 in oesophageal carcinoma.

AIM: To examine the expression of HLA-ABC and HLA-DR major histocompatibility (MHC) antigens and intercellular adhesion molecule (ICAM)-1 in normal, inflamed, metaplastic, and neoplastic oesophageal tissue and in freshly disaggregated tumours. METHODS: Sequential sections of frozen tissue and cytospins of freshly disaggregated tumour were stained using the ABC peroxidase system and monoclonal antibodies specific for HLA-ABC, HLA-DR and ICAM-1. RESULTS: Normal oesophageal tissue showed positive staining for HLA-ABC in the basal layers of the oesophageal squamous epithelium and on the epithelial cells of the submucosal oesophageal glands. HLA-DR and ICAM-1 were not detected in either of these cell types. In 20 of 37 (54%) carcinomas HLA-ABC was expressed weakly, with heterogeneous expression in nine (24%). Two tumours showed strong expression of HLA-ABC, but 15 of 37 (41%) were negative. HLA-DR and ICAM-1 were expressed weakly in six of 37 (16%) carcinomas without correlation with each other or with the expression of HLA-ABC. CONCLUSIONS: HLA-ABC is absent from a high proportion of oesophageal carcinomas (41%) and is otherwise variably and weakly expressed with strong expression in only a small fraction (3%). In other carcinomas there is a higher level of HLA-ABC expression. This discrepancy may partly explain the aggressive nature of oesophageal carcinomas. HLA-DR and ICAM-1 are not normally expressed on those cells from which oesophageal carcinomas are thought to arise. The limited expression found here could suggest a partial or inhibited immune response against oesophageal carcinoma. In vivo repressive factors may be involved.

Lymphocyte infiltration in oesophageal carcinoma: lack of correlation with MHC antigens, ICAM-1, and tumour stage and grade.

Infiltration by T lymphocytes into oesophageal carcinomas was assessed immunohistochemically, total T lymphocyte numbers by staining for CD3 and activated T lymphocytes by staining for CD25. Five squamous carcinomas and seven adenocarcinomas, resected without neoadjuvant treatment, were studied. Computer aided quantitation showed that total numbers of tumour infiltrating CD3 positive cells were highly variable (range 48-1673 cells/mm2). They were located largely in the stromal (87.9-99.2%) rather than intratumoral regions. Up to 84% of tumour infiltrating T lymphocytes were CD25 positive, although the median figure was 33%. There was no correlation between T lymphocyte infiltration or activation and expression of class I and II histocompatibility antigens, intercellular adhesion molecule-1, tumour stage or grade. These results imply that the local inflammatory response in oesophageal carcinomas is deregulated, which may be a factor contributing to the aggressive nature of the tumours.

Clinical significance of selective decline of donor-reactive IL-2-producing T lymphocytes after renal transplantation.

Limiting dilution analysis technique was used to enumerate the circulating precursor frequency of donor and third-party-reactive helper T lymphocytes (HTLpf) in 28 renal allograft recipients before (pre-tx) and at three intervals (T1: 60-90 days, T2: 120-180 days, T3: 360-1620 days) after transplantation (post-tx). Two patterns of responses were identified, in group 1 (n = 12), a five to 31-fold reduction of donor-reactive HTLpf (ranging from 1/19231-1/62500) occurred within 90-1620 days post-tx, while in group 2(n = 16), no significant changes of donor-reactive HTLpf were seen. In both groups, the third-party-reactive HTLpf in most of these patients remained largely unchanged throughout the study period. The number of HLA-DR mismatches, total number of rejection episodes, serum creatinine levels, and biopsy findings at T3 were compared in both groups using Fisher's exact probability, and the Mann-Whitney test. We found that 11 patients (92%) in group 1 were HLA-DR compatible with donors, while nine (56%) patients in group 2 were HLA-DR compatible with donors, p = 0.04. In group 1 eight rejection episodes occurred in five (41.6%) patients during the study period, compared to 33 in 13 (81%) patients in group 2, p = 0.03. Group 1 had a significantly lower serum creatinine level (at T3); median: 136 vs 165 mumol/l for group 2, p = 0.03. Biopsy indicated no rejection (at T3) in eight (66%) patients in group 1 as compared to three (18%) patients in group 2, p = 0.03. Taken together, these results indicate that the frequency of circulating HTLpf correlate with the clinical status of the graft. Therefore monitoring of HTLpf in the peripheral blood could be useful in predicting graft outcome and selecting patients for reducing immunosuppression.

A cluster of single nucleotide polymorphisms in the 5'-leader of the human dopamine D3 receptor gene (DRD3) and its relationship to schizophrenia.

The association between schizophrenia and the Ser9Gly variant of the dopamine D3 receptor gene (DRD3) has been the subject of numerous studies. Under meta-analysis this site, or one or more in linkage disequilibrium with it, appears to contribute a small increase to the relative risk of schizophrenia. In this study, 768 bp of the 5'-leader region of DRD3 mRNA was screened for polymorphisms to assess their contribution to the association of DRD3 with schizophrenia. A cluster of three single nucleotide polymorphisms (SNPs) was identified in tight linkage disequilibrium with each other and with the Ser9Gly polymorphism. One of the 5'-leader SNPs encodes a Lys9Glu variant within a 36 amino acid residue stretch of an upstream open reading frame (uORF). Two common haplotypes are found in the population examined; one is linked to the Ser9 coding variant and the other to the Gly9 variant. A panel of 73 schizophrenic patients and 56 matched controls recruited from the East Anglia region of the United Kingdom was screened for disease association at these sites. Since the 5'-leader and coding sites are in tight disequilibrium, the combined genotype of all 4 sites was scored for each patient. A significant association was seen between disease and the frequency distribution of these genotypes (chi2 = 13.19, d.f. = 3, P = 0.0042; Cochran method for sparse cells applied). A 20% excess of one of the heterozygous genotypes, in which the sequences differ at three of the four SNPs, including Ser9/Gly9 in the receptor and Lys9/Glu9 in the uORF, was found in the patient group. An absence of association of disease with the Ser9Gly polymorphism had previously been reported for this panel. This suggests that these SNPs and the corresponding coding changes may exert a combined or synergistic effect on susceptibility to schizophrenia.

The exfoliation of free malignant cells in the peritoneal cavity during resection of colorectal cancer.

The presence of free malignant cells in the peritoneal cavity following potentially curative resection of colorectal cancer may be responsible for some cases of local recurrence. Twenty-six patients (age 58-88 years, male:female 12:14) undergoing colorectal cancer surgery were studied to determine frequency of occurrence of tumour cells in the peritoneal cavity using an immuno-magnetic cell separation technique. Peritoneal washings were collected after opening the peritoneal cavity and also prior to closure. A murine monoclonal antibody specific for epithelial cells was added to the peritoneal washings followed by magnetic labelled goat-antimouse antibody. The labelled tumour cells were retrieved by passing through the magnetic activated cell sorter (MACS). Tumour cells were detected in peritoneal washings from 11 patients (preresection only n = 0, post-resection only n = 6, and both n = 5). Five of these 11 patients had potentially curative surgery. This study supports the use of MACS in detecting free malignant cells in the peritoneal cavity.

Telomerase activity and patient survival after surgery for gastric and oesophageal cancer.

OBJECTIVE: The ribonucleoprotein telomerase extends telomeres in cancer cells and has been proposed as a prognostic marker for cancer. We measured telomerase expression in proximal adenocarcinomas (those arising in the distal oesophagus or at the gastro-oesophageal junction) and distal adenocarcinomas (those arising in the corpus or antrum of the stomach) of the foregut, and correlated telomerase activity with pathological stage and post-operative survival. DESIGN: Surgical specimens were collected from patients undergoing resections for gastric and oesophageal carcinomas. Haematoxylin and eosin histology provided data on the pathological tumour stage and pathological node stage. METHODS: The telomerase activity of cancer specimens was determined using the telomeric repeat amplification protocol. A single pathologist, blinded to the results of the telomerase assays, reviewed all slides of cancers to assign T and N stages. RESULTS: The cancers exhibited a wide range of telomerase expression. There was no significant difference between the telomerase activity of proximal adenocarcinomas (median, 551 U; 95% confidence interval, 154-2394 U; n = 26) and distal adenocarcinomas (median, 703 U; 95% confidence interval, 139-1618 U; n = 20). Distal adenocarcinomas expressing high telomerase activity (greater than the median) were significantly more advanced with regard to T stage than distal cancers expressing low telomerase levels (less than the median; P = 0.03, Mann-Whitney test). In distal adenocarcinomas, high telomerase activity was associated with poor patient survival (median 3 months) compared to low telomerase activity (median survival 22.4 months; P = 0.01, log-rank test). No such differences were observed for proximal adenocarcinomas. CONCLUSIONS: There is a difference between gastric and oesophageal/gastro-oesophageal junction adenocarcinomas in terms of the relationship with telomerase expression and clinico-pathological variables. Among patients with distal gastric adenocarcinoma, telomerase activity correlates with markers of advanced disease, whereas this relationship does not hold true in oesophageal/gastro-oesophageal junction adenocarcinomas. Telomerase activation may occur at different stages of the formation of the malignant phenotype in these two cancers and may reflect differences in their pathogenesis. Telomerase could be a prognostic marker in gastric but not in oesophageal adenocarcinoma.

Changes in mandibular condyle morphology related to tooth wear in a prehistoric human population.

The exact relationship between tooth wear and changes in the shape of the articulating surface of the mandibular condyles is unknown. It is assumed that to some degree these changes are in response to the loads on the joint generated during chewing. Excavated skeletal remains from sites of primitive human habitation provide an opportunity to study the masticatory system of human beings in their natural context, with the features of form and function expressed to their full potential. Accurate measurements of the condylar head can also be made on such remains. A collection of skulls from an excavation site known as Oakhurst on the South African coast provided material for this study. Occlusal wear was measured using the scale devised by Molnar; approximal wear was assessed by measuring the size of the wear facet between the first and second molars. The width of the condyles were measured directly, and a method was devised to measure changes in the proportion of medial and lateral articulating facets. An increase in the size of the lateral facet of the condyle was found to be associated with increased tooth wear. It is suggested that the direction of condylar remodelling may give an indication of the reaction at the joint to optimal mastication.