p185HER2 overexpression in human breast cancer using molecular and immunohistochemical methods.

With the successful clinical trials of the engineered antibody Herceptin (in advanced-stage breast cancer) and adriamycin-based chemotherapy regimens (in the adjuvant setting), the need to detect p185HER2 overexpression or associated amplification of the coding gene HER2 in breast cancer patients is escalating. Twenty to 30% of breast carcinomas have overexpression of p185HER2. This condition correlates with poor patient prognosis and predicts response to chemotherapy in lymph node-positive patients.

The role of genomic instability in human carcinogenesis.

Neoplastic cells typically possess numerous genomic lesions, which may include sequence alterations (point mutations, small deletions, and insertions) and/or gross structural abnormalities in one or more chromosomes (large-scale deletions, rearrangements, gene amplifications). Based upon this general observation, it has been suggested that cancer cells are genetically unstable, and that acquisition of genomic instability may represent an early step in the process of carcinogenesis and a general feature of many human tumors. Numerous studies have appeared that characterize the nature and frequency of occurrence of various molecular lesions in human tumors, and significant progress has been made towards the elucidation of the molecular mechanisms that govern genetic stability in normal cells and genetic instability in neoplastic cells.

Polymorphisms in the genes for coagulation factors II, V, and VII in patients with ischemic heart disease.

Background: Cardiovascular disease remains the leading cause of mortality in the United States, accounting for approximately 33% of all deaths in this country. Of these deaths, most are due to acute myocardial infarctions (AMIs), which are associated with thrombotic coronary artery obstruction and/or occlusion. These events could potentially be due to alterations in genes coding for coagulation factors.

Rapid detection of HSV from cytologic specimens collected into ThinPrep fixative.

Objective: Herpes simplex virus (HSV) infection is associated with substantial morbidity and mortality in neonates. A diagnosis of HSV on cervical cytologic studies could lead to a cesarean section, with an increase in the risk of maternal morbidity. The identification of viral lesions in sexually active women has medical and social implications.

Prevalence of the C282Y and H63D polymorphisms in a multi-ethnic control population.

Recently a candidate gene for hereditary hemo-chromatosis, HFE, was identified. The finding raises the possibility for genetic testing to provide earlier detection and more complete genotypic evaluation of hemochromatosis affected individuals. We determined the frequency of the HFE polymorphisms, C282Y and H63D, in a randomly selected multi-ethnic control population for establishment of a hemochromatosis genetic testing program.

The use of monoclonal antibody R92F6 and polymerase chain reaction to confirm the presence of parvovirus B19 in bone marrow specimens of patients with acquired immunodeficiency syndrome.

Background: Parvovirus B19 infection is a cause of chronic anemia and red cell aplasia in patients with acquired immunodeficiency syndrome (AIDS) and in other immunocompromised hosts. Anemia in AIDS patients has a multifactorial etiology, with parvovirus B19 infection being an infrequent but nevertheless treatable cause. Therapy with intravenous immune globulin can result in rapid improvement of parvovirus-induced anemia.

Homocysteine screening of a female Hispanic population.

A total of 821 women of Hispanic descent aged 21-65 years, were screened for total and high density lipoprotein (HDL) cholesterol through outpatient clinics and public screening. Of this group, 78 were invited back for further testing because they had a total cholesterol/HDL cholesterol ratio exceeding 4.5 indicative of high risk for cardiovascular disease.

Applications of forensic identity testing in the clinical laboratory.

DNA analysis is becoming routine in the clinical laboratory for the diagnosis of human diseases using various tissue sources. Most clinical specimens are followed by tracking forms that include patient demographic data, accession number, and date and time of collection. As part of a thorough quality assurance program, proper documentation of test requisitions and tracking forms is mandatory.

Molecular characterization of the myxosporean associated with parasitic encephalitis of farmed Atlantic salmon Salmo salar in Ireland.

During seasonal epizootics of neurologic disease and mass mortality in the summers of 1992, 1993 and 1994 on a sea-farm in Ireland, Atlantic salmon Salmo salar smolts suffered from encephalitis associated with infection by a neurotropic parasite. Based on ultrastructural studies, this neurotropic parasite was identified as an intercellular presporogonic multicellular developmental stage of a histozoic myxosporean, possibly a Myxobolus species. In order to generate sequence data for phylogenetic comparisons to substantiate the present morphological identification of this myxosporean in the absence of detectable sporogony, polymerase chain reaction (PCR), Southern blot hybridization, dideoxynucleotide chain-termination DNA sequencing, and in situ hybridization (ISH) were used in concert to characterize segments of the small subunit ribosomal RNA (SSU rRNA) gene.

Proliferative fraction, bcl-1 gene translocation, and p53 mutation status as markers in mantle cell lymphoma.

The molecular genetic hallmark of mantle cell lymphomas (MCL) is the reciprocal translocation t(11;14)(q13;q32) which juxtaposes the bcl-1 proto-oncogene to one of the joining segments of the immunoglobulin heavy chain gene. This translocation is very common in MCL and occurs in up to 70% of these malignancies. Due to the aggressive nature of MCL, markers identifying tumor progression and clinical outcomes are necessary.

Rapid multiplex analysis for the factor V Leiden and prothrombin G20210A mutations associated with hereditary thrombophilia.

Thromboembolic episodes are common events and affect approximately one in 1,000 persons annually. Pulmonary embolism alone accounts for 50,000 to 100,000 deaths per year in the United States with > 50% of those being elderly persons. Resistance to activated protein C is the most common inherited disorder associated with hereditary thrombophilia.

The effect of pathologic substances and adulterants on the DNA typing of urine.

Human urine has not been adequately investigated as a potential source of DNA for forensic identity testing. The advent of polymerase chain reaction technology has made possible the analysis of previously undetectable levels of nucleic acids from human urine and other body fluids lacking nucleated cells. In this study, we evaluated the ability to genotype DNA extracted from adulterated urine specimens using the AmpliType PM + DQA1 PCR amplification and typing system.

Double heterozygosity for mutations in the BRCA1 and BRCA2 genes in a breast cancer patient.

Background: Germline mutations in the tumor suppressor genes BRCA1 and BRCA2 confer substantial increased lifetime risk for breast cancer, and in the case of BRCA1, for ovarian carcinoma as well. These two genes alone account for the vast majority of hereditary breast cancer families. Numerous mutations have been described in each gene, the majority of which are small insertions or deletions resulting in expression of a truncated protein.

Myocardial cell death and apoptosis in hibernating myocardium.

Objectives: This study was designed to study apoptosis in hypoperfused hibernating myocardium subtending severe coronary stenosis.

Background: Apoptosis contributes to myocyte death in acute myocardial infarction.

Methods: A left anterior descending coronary artery stenosis was created in 13 pigs and maintained for 24 h (n = 4), 7 days (n = 5) and 4 weeks (n = 4) to reduce coronary blood flow by a mean of 34% with severe regional myocardial systolic dysfunction, as documented by echocardiography.

Correlation between morphologic and nonmorphologic prognostic markers of neuroblastoma.

Morphologic (Shimada classification--SC, original and modified histologic grades--OHG and MHG) and nonmorphologic (serum LDH, 1 p del, DNA index, N-myc copy number, telomerase activity, and expression of MRP, MDR1, and TRK) prognostic markers for NB have been reviewed. The functional role of these nonmorphologic markers in the development and progression of this disease include abnormal cell proliferation, resistance to chemotherapeutic agents, and induction of apoptosis. A statistically significant association between high OHG/MHG (grade 3), DNA index of 1 (diploidy), > 1 copy of N-myc per haploid genome and serum LDH of > or = 1500 IU/1 (p < 0.

Molecular diagnosis of B- and T-cell lymphomas: fundamental principles and clinical applications.

Molecular diagnostic assays have become routine in the evaluation of lymphoid malignancies. Both Southern transfer and polymerase chain reaction (PCR) technologies are used to assess for B- and T-cell clonality, the presence of rearrangements involving protooncogenes such as bcl-1 and bcl-2, and the monitoring of minimal residual disease. We review the fundamentals of B- and T-cell ontogeny as well as the basic principles of the Southern transfer and PCR assays and their applications to the diagnosis of lymphoid malignancies.

Life threatening pulmonary embolus in a factor V Leiden carrier on oral contraceptives: a case report.

Venous thromboembolism is a serious, potentially lethal health problem affecting one per 1,000 people annually. Major surgery, the use of oral contraceptives, complicated pregnancy, fractures, and immobilization increase the risk of thrombosis. In addition to these factors, thrombosis is associated with inherited deficiencies of antithrombin III, protein C, and protein S.

Application of forensic identity testing in a clinical setting. Specimen identification.

Specimen identification is a carefully controlled factor in clinical laboratory testing. However, on occasion, despite surmountable efforts to prevent misidentification, a specimen is either mislabeled or an identifier is lost. Recently, we experienced a case of questionable mix-up of surgical specimens where the surgeon and patient questioned the biopsy site and size of specimen as indicated in the anatomic pathology report.

Correction of the DNA repair defect in Fanconi anemia complementation groups A and D cells.

We have previously isolated from Fanconi anemia, complementation groups A (FA-A) and D (FA-D) cells, a DNA endonuclease complex which is defective in its ability to incise DNA containing interstrand cross-links produced by psoralen plus UVA light. The repair capabilities of the FA complexes, compared with those of the corresponding normal complex, have now been examined using two types of complementation analysis. First, introduction of the normal complex, by electroporation, into 8-methoxypsoralen (8-MOP) plus UVA treated FA-A and FA-D cells resulted in correction of their repair defect, determined by measuring repair-related unscheduled DNA synthesis (UDS).

Identification of urine specimen donors by the PM+DQA1 amplification and typing kit.

We evaluated the ability to genotype DNA extracted from urine samples, which were previously submitted for toxicological analysis, by either the AmpliType HLA DQ alpha or the combined PM+DQA1 amplification and typing systems. Initial experiments were conducted on fresh urine, which was either processed fresh or frozen for one week at -20 degrees C, from male and female volunteers. Although male urine is noted for containing minimal numbers of nucleated cells when compared with female urine, we were able to type these samples without difficulty.

The underlying molecular mechanism of apolipoprotein E polymorphism: relationships to lipid disorders, cardiovascular disease, and Alzheimer's disease.

Apolipoprotein E (apo E) polymorphism has important clinical correlates, including disorders of lipoprotein metabolism and atherosclerosis. This article provides a detailed methodology for apo E genotyping and discusses the link between apo E genotype and type III hyperlipoproteinemia, coronary heart disease (CHD), stroke, and Alzheimer's disease (AD). Although apo E genotype appears to provide significant information concerning the genetic component of CHD and AD risk, more research is needed before genotyping can be recommended as a routine screening tool.