Loss of heterozygosity at 11q22 correlates with low progesterone receptor content in epithelial ovarian cancer.

Forty-seven epithelial ovarian cancers were analyzed for loss of heterozygosity (LOH) at D11S35 (11q22), close to the progesterone receptor (PR) gene, and for tumoral estrogen receptor (ER) and PR content. Thirty-eight of 47 tumors were informative, and, of these, 14 exhibited LOH. There was a significant association (P = 0.014) between D11S35 LOH and low tumoral PR content. For all informative tumors, there was no correlation between ER and PR; however, exclusion of tumors with LOH from the informative series revealed a linear correlation between tumoral ER and PR (P = 0.013), and established ER (P = 0.025) and PR (P = 0.05) content as significant factors in relation to patient survival. Patients with ER-rich tumors with D11S35 LOH had particularly poor survival compared with ER-rich, D11S35 heterozygous, no loss patients (P = 0.014). Analysis of the same tumors using two other microsatellites, D11S935 (11p13) and NM23 (17q22), showed no statistically significant relationships, although there were nonsignificant trends for the correlation of ER and PR expression in informative tumors without allele loss at these loci. We propose that genomic structural alteration at or close to the PR gene locus has biological and clinical sequelae in ovarian cancer.

A reliable method for the analysis of human MHC class II alpha and beta chains by Western blotting of IEF gels.

The solubility and ensuing transfer problems associated with IEF separation and blotting of membrane proteins have been eliminated by confining NP40 to the sample region of the gel and replacing it with octylglycoside in the separating gel. This permitted excellent separation of both the alpha and beta chains of the human MHC antigens and their subsequent detection by Western blotting.

Methods of detecting mature human Ia-like antigen and their metabolic precursors.

Ia antigens were shown to be present in the cell almost exclusively as mature alpha beta dimers which split into separate alpha and beta chains after boiling in SDS. In contrast metabolically labelling the cells with [35S]methionine resulted in only free alpha and beta chains being labelled. It is concluded that this widely used type of labelling, although useful for studying intermediate synthesis, should not be used for labelling mature cell surface molecules.

The use of detergents in velocity sedimentation of cell culture IgM.

The biosynthetically radiolabelled secreted proteins of human lymphoblastoid cell lines have been analysed by sucrose gradient velocity sedimentation. The addition of detergent to the gradient buffer dramatically improves the recovery of immunoglobulins from cell line supernatants, allowing the gradient fractions to be analysed further both serologically and biochemically. The results of these analyses show that IgM is synthesised in the 19S form and can be clearly separated from other components which include free light chains. Immunoglobulin appears to form a much larger portion of the proteins secreted by human lymphoblastoid lines than has formerly been recognised, a finding which may be of practical importance for the development and exploitation of human monoclonal antibodies.

A mild procedure for separating polypeptide chains prior to immunoprecipitation and western blotting analysis.

Conventional cleavage of linked polypeptide chains by heating in SDS can so alter molecular structure as to interfere with antibody binding, on which both immunoprecipitation and 'western blotting' depend. As an alternative, gentle treatment with acid at room temperature or at 0 degrees C was effective in separating the alpha and beta chains of human MHC Class II glycoprotein dimers and proved superior in terms of preservation of at least one labile epitope on the beta chain.

A human lymphoblastoid B line with an abnormally large MHC class II beta chain.

We have examined the MHC class II beta chains in lymphoblastoid cell lines from over 200 individuals and describe one line which possesses, in addition to normal beta chains, a species of beta chain of unusually high Mr and abnormal pI which appears to be a product of the DR locus. This abnormality in Mr, detected by SDS-gel electrophoresis, was apparent only in the presence of mercaptoethanol and was shown to be due to difference in polypeptide chain length rather than to extra glycosylation.

Detection of Ia epitopes by monoclonal antibody blotting.

Monoclonal antibodies directed against human Ia alpha- and beta-subunit chains have been used as probes to detect polypeptides carrying recognized antigenic determinants or epitopes following two-dimensional PAGE separation. Approximately 4 sets of components differing in isoelectric point but not in molecular weight are recognized by each antibody. The anti-alpha-chain, McAb, reacts weakly with spots designated as epsilon but neither antibody recognizes Im, Ii or delta determinants under the conditions tested.

Southern blot analysis of DNA extracted from paraffin wax-embedded tissue.

Genetic analysis of multi-generation families using RFLP is often incomplete because fresh tissue is not available for the isolation of DNA. However, in many instances archival material exists, usually formalin-fixed in paraffin blocks. Procedures have been described for extracting DNA from such specimens. Many parameters affect the quality of the DNA eventually extracted and this in turn determines whether the DNA can be successfully digested with restriction enzymes and probed (Dubeau et al., 1986; Warford et al., 1988). In essence, the procedures followed at the time of fixation largely determine whether archival material will yield useful results. We have re-examined the effect of varying the fixation time but, more importantly, we have assessed the validity of RFLP results obtained from fixed tissue.

Ethnicity, maternal risk, and birth weight among Hispanics in Massachusetts, 1987-89.

National data reveal that low birth weight and infant mortality rates among Hispanics are, in general, between the rates for whites and those for blacks. The question remains, do differences in low birth weight reflect distributions of known risk factors, or do ethnic differences persist after simultaneously adjusting for intervening variables? In this study, Massachusetts birth certificate data for 206,973 white non-Hispanic infants and 19,571 Hispanic infants are used to examine differences in low birth weight between white non-Hispanic and Hispanic infants, as well as variation among seven subgroups of Hispanic mothers--Puerto Rican, Dominican, Central American, South American, Mexican, Cuban, and other Hispanic. Regression analysis is used to estimate the association between risk factors and birth weight and the relative risk of low birth weight. Risk factors include ethnicity, demographic characteristics, biological factors, access to prenatal care, and infants' conditions. Results indicate substantial variation in mean birth weight, low birth weight, and levels of risk among Hispanic subgroups and between Hispanics and white non-Hispanics. Puerto Rican infants had the lowest mean birth weight and, in general, the highest level of risk factors in this population. None of the adjusted odds ratios for low birth weight for any Hispanic group was significantly elevated at the 95 percent level compared with white non-Hispanics. Findings in this study confirm the previous observations of the wide variation among Hispanic subgroups and the high level of risk among Puerto Ricans. Results of this study also raise some interesting questions about the differential relationship between ethnicity and birth weight, ethnicity and low birth weight, and the significance of maternal place of birth as a proxy measure of adaptation or acculturation.

Overview of causes and costs of injuries in Massachusetts: a methodology for analysis of state data.

Massachusetts has developed the first State profile of the causes and costs of injury based on the national study, "Cost of Injury in the United States: A Report to Congress." Incidence of fatal injuries is based on Massachusetts data; nonfatal hospitalized injuries, on Massachusetts age and sex rates and U.S. cause data; and nonhospitalized injuries, on U.S. rates applied to Massachusetts census data. Lifetime costs per injured person are based on national data adjusted for higher personal health care expenditures and for higher mean annual earnings in Massachusetts. The estimated total lifetime cost for the 1.4 million injuries that occurred in 1989 is $4.4 billion--$1.7 billion for health care and $2.7 billion for lost earnings. Injuries attributed to motor vehicles and falls account for more than half of the total cost. The other cause categories are poisonings, fire-burns, firearms, drowings-near drownings, and other. For every person who dies from an injury, 17 people are hospitalized, and an estimated 535 people require outpatient treatment, consultation, or restricted activity. Development of a State-based cost report can be useful in monitoring the contribution of injuries to health status and in planning effective injury prevention strategies in a community-based health care system. The methodology described in this paper can be replicated by other States through accessing their State-specific mortality and hospital discharge data bases.

A significant proportion of patients with osteosarcoma may belong to Li-Fraumeni cancer families.

We studied the pedigrees of 17 index patients with osteosarcoma, recording malignant disease and cause of death for first- and second-degree relatives. There were seven cancers and five cancer deaths per 2151.5 person-years in first-degree relatives of osteosarcoma patients under the age of 50 years, a significantly greater incidence than in an age- and sex-matched population group (p < 0.001). This excess of malignancy was largely due to two families which fulfilled the criteria for the Li-Fraumeni cancer family syndrome. Both of these families were shown to have the genetic alterations in the p53 gene which have been implicated in this syndrome. Our study suggests that orthopaedic surgeons seeing new cases of osteosarcoma should arrange screening for familial malignancy.

Maternal ethnicity and birthweight among blacks.

This paper examines the association of ethnicity and birthweight, adjusted for other maternal and infant characteristics, among black women who gave birth in Massachusetts from 1987 through 1989. Data are drawn from the standard certificate of live birth, which includes questions on race and ethnicity/ancestry as well as birthweight; maternal sociodemographic and biological characteristics; access to prenatal care; and infant characteristics. The study cohort consists of 18,571 black infants and a comparison group of 206,358 non-Hispanic white infants. Infants whose mothers reported their race as black were further categorized into six ethnic groups: American, Haitian, West Indian, Cape Verdean, Hispanic, and other black. In addition to descriptive analyses, we used multiple linear regression to measure the association between ethnicity, other characteristics, and birthweight; and we used multiple logistic regression to measure the odds ratio of low birthweight (ranging from 500 g to 2499 g) for the six black ethnic groups, adjusted for other characteristics. Results indicate that Americans have lower mean birthweight and generally higher levels of risk than other black ethnic groups. Compared to the reference group of non-Hispanic whites, Americans (OR = 1.49), other blacks (OR = 1.41), and West Indians (OR = 1.37) have significantly elevated relative risks of low birthweight.

The process and outcome of hospital care for Medicaid versus privately insured hospital patients.

Some have argued that low Medicaid payment rates compromise the accessibility and quality of medical care for Medicaid beneficiaries. In this study we compare the process and outcome of hospital care for Medicaid versus privately insured hospital patients. We studied 4,033 emergency patients admitted with a principal diagnosis of acute myocardial infarction, to Massachusetts hospitals in 1987. After we statistically adjusted for differences among patients relating to clinical and demographic characteristics and the type of hospital where treatment occurred, we found that the Medicaid patients had longer hospital stays but were less likely to receive three selected coronary procedures. Moreover, after controlling for confounding variables, we found the risk of death for Medicaid patients to be almost twice as high as for privately insured patients.

Inequities in hospital care, the Massachusetts experience.

This study examined the relationship between patient insurance status and the process and outcome of hospital care in Massachusetts, a state that has had an uncompensated care pool for paying hospitals since 1986. This study examined data on 4,972 patients admitted to a Massachusetts hospital on an emergency basis in 1987 and diagnosed with acute myocardial infarction. We classified these patients into three groups: having fee-for-service insurance, having prepaid coverage through a health maintenance organization (HMO), or being uninsured at the time of hospital admission. Results showed treatment differences by insurance status and significantly greater mortality rates for uninsured patients than for either fee-for-service or HMO patients. Our findings indicate that in Massachusetts the process and outcome of hospital care do differ by insurance status.

Ribonuclease inhibitors from mouse ascites cells.

Ribonuclease inhibitors were found to be present attached to mouse ascites ribosomes and in the post-ribosomal supernatant. Both inhibitors inhibited pancreatic RNAses A and B and two endonucleases prepared from ascites cells but did not inhibit RNAses T1 or N1. Both inhibitors had the same sedimentation coefficient and this taken with the results above suggest that they are identical. The inhibitor was shown to interact directly with the RNAse itself.