Genomic analyses of germline and somatic variation in high-grade serous ovarian cancer.

BACKGROUND: High-grade serous ovarian cancers (HGSCs) display a high degree of complex genetic alterations. In this study, we identified germline and somatic genetic alterations in HGSC and their association with relapse-free and overall survival. Using a targeted capture of 557 genes involved in DNA damage response and PI3K/AKT/mTOR pathways, we conducted next-generation sequencing of DNA from matched blood and tumor tissue from 71 HGSC participants. In addition, we performed the OncoScan assay on tumor DNA from 61 participants to examine somatic copy number alterations (SCNA). RESULTS: Approximately one-third of tumors had loss-of-function (LOF) germline (18/71, 25.4%) or somatic (7/71, 9.9%) variants in the DNA homologous recombination repair pathway genes BRCA1, BRCA2, CHEK2, MRE11A, BLM, and PALB2. LOF germline variants also were identified in other Fanconi anemia genes and in MAPK and PI3K/AKT/mTOR pathway genes. Most tumors harbored somatic TP53 variants (65/71, 91.5%). Using the OncoScan assay on tumor DNA from 61 participants, we identified focal homozygous deletions in BRCA1, BRCA2, MAP2K4, PTEN, RB1, SLX4, STK11, CREBBP, and NF1. In total, 38% (27/71) of HGSC patients harbored pathogenic variants in DNA homologous recombination repair genes. For patients with multiple tissues from the primary debulking or from multiple surgeries, the somatic mutations were maintained with few newly acquired point mutations suggesting that tumor evolution was not through somatic mutations. There was a significant association of LOF variants in homologous recombination repair pathway genes and high-amplitude somatic copy number alterations. Using GISTIC analysis, we identified NOTCH3, ZNF536, and PIK3R2 in these regions that were significantly associated with an increase in cancer recurrence and a reduction in overall survival. CONCLUSIONS: From 71 patients with HGCS, we performed targeted germline and tumor sequencing and provided a comprehensive analysis of these 557 genes. We identified germline and somatic genetic alterations including somatic copy number alterations and analyzed their associations with relapse-free and overall survival. This single-site long-term follow-up study provides additional information on genetic alterations related to occurrence and outcome of HGSC. Our findings suggest that targeted treatments based on both variant and SCNA profile potentially could improve relapse-free and overall survival.

Genomic Analyses of Germline and Somatic Variation in High-Grade Serous Ovarian Cancer.

Background High-grade serous ovarian cancers (HGSCs) display a high degree of complex genetic alterations. In this study, we identified germline and somatic genetic alterations in HGSC and their association with relapse-free and overall survival. Using a targeted capture of 577 genes involved in DNA damage response and PI3K/AKT/mTOR pathways, we conducted next-generation sequencing of DNA from matched blood and tumor tissue from 71 HGSC participants. In addition, we performed the OncoScan assay on tumor DNA from 61 participants to examine somatic copy number alterations. Results Approximately one-third of tumors had loss-of-function germline (18/71, 25.4%) or somatic (7/71, 9.9%) variants in the DNA homologous recombination repair pathway genes BRCA1, BRCA2, CHEK2, MRE11A, BLM , and PALB2 . Loss-of-function germline variants also were identified in other Fanconi anemia genes and in MAPK and PI3K/AKT/mTOR pathway genes. Most tumors harbored somatic TP53 variants (65/71, 91.5%). Using the OncoScan assay on tumor DNA from 61 participants, we identified focal homozygous deletions in BRCA1, BRCA2, MAP2K4, PTEN, RB1, SLX4, STK11, CREBBP , and NF1 . In total, 38% (27/71) of HGSC patients harbored pathogenic variants in DNA homologous recombination repair genes. For patients with multiple tissues from the primary debulking or from multiple surgeries, the somatic mutations were maintained with few newly acquired point mutations suggesting that tumor evolution was not through somatic mutations. There was a significant association of loss-of-function variants in homologous recombination repair pathway genes and high-amplitude somatic copy number alterations. Using GISTIC analysis, we identified NOTCH3, ZNF536 , and PIK3R2 in these regions that were significantly associated with an increase in cancer recurrence and a reduction in overall survival. Conclusions From 71 patients with HGCS, we performed targeted germline and tumor sequencing and provided a comprehensive analysis of these 577 genes. We identified germline and somatic genetic alterations including somatic copy number alterations and analyzed their associations with relapse-free and overall survival. This single-site long-term follow-up study provides additional information on genetic alterations related to occurrence and outcome of HGSC. Our findings suggest that targeted treatments based on both variant and SCNA profile potentially could improve relapse-free and overall survival.

Morphological, electrical & antibacterial properties of trilayered Cs/PAA/PPy bionanocomposites hydrogel based on Fe3O4-NPs.

Bionanocomposites hydrogel based on conducting polymers were successfully fabricated from chitosan/polyacrylic acid/polypyrrole (CS/PAA/PPy) as well as the magnetite nanoparticle (Fe3O4-NPs) was prepared via co-precipitation method. In addition, different ratios of Fe3O4-NPs were added to the prepared bionanocomposites to enhance the antimicrobial and the electrical conductivity of the prepared conductive hydrogel. Furthermore, the morphology, the swelling percent, antimicrobial activity and the dielectric properties of the prepared conducting bionanocomposites hydrogel were investigated. The antibacterial activities of the experienced microbes were improved with the increasing the loading of Fe3O4-NPs in conducting Bio-nanocomposites hydrogel. Moreover, the DC-conductivity was examined and our resulted indicated that the DC-conductivity was enhanced by increasing the loadings of Fe3O4-NPs compared to that of the pure CS/PAA as well as CS/PAA/PPy.

Laparoscopic donor nephrectomy: single-center experience in Egypt with 400 consecutive cases.

INTRODUCTION: In this study, we present our experience with laparoscopic donor nephrectomy and evaluate the outcomes of donors and recipients. PATIENTS AND METHODS: Between March 2003 and August 2006, 400 laparoscopic donor nephrectomies were performed in our institution. Donors were evaluated for renal vasculature using computed tomography angiography. We used the left kidney in 329 donors and the right kidney in 71. Donor surgeries were done transperitoneally using three trocars on the left side and four trocars on the right side. Kidneys were extracted manually through a 7-cm Pfanenstiel incision. RESULTS: All cases were completed laparoscopically. Mean operative time was 117 +/- 34 minutes. Mean blood loss was 56 +/- 28 mL. None of the donors required a blood transfusion. Mean warm ischemia time was 2.6 +/- 0.4 minutes. The mean renal artery length was 3.1 +/- 0.4 cm; the mean renal vein length was 2.4 +/- 1.2 cm. Mean hospital stay was 2.1 days. No donor required readmission. Kidneys were transplanted successfully and the mean recipient creatinine on discharge was 1.2 +/- 0.6 mg/dL. One patient had a renal artery thrombosis on postoperative day 2. Another patient with double renal arteries had thrombosis of the smaller artery just after surgery. Acute tubular necrosis was seen in 17 patients, four of whom required dialysis. Kidney function recovered thereafter in all acute tubular necrosis cases. CONCLUSION: Laparoscopic surgery is a minimally invasive approach for living donor nephrectomy with good functional outcomes. The donor benefits from lesser morbidity without compromising the anatomic or physiological outcome of the nephrectomized kidney.

Laparoscopic donor nephrectomy for pediatric recipient.

INTRODUCTION: To present our outcome of laparoscopic donor nephrectomy for pediatric recipients, who may pose special challenges. MATERIALS AND METHODS: Since March 2003, we performed more than 400 laparoscopic donor nephrectomies for 39 pediatric recipients (age less than 17 years of age). The preoperative, intraoperative, and postoperative data were reviewed to analyze the outcomes of these cases. We used the left kidney in 26 and the right kidney in 13 cases. Seven cases had double renal arteries, which were reconstructed on the bench. RESULTS: The mean donor and recipient ages were 31 +/- 5 years and 13 +/- 4 years, respectively. The mean donor operative time was 2.1 hours (range 1.2 to 3.2). The warm ischemia time averaged 3 +/- 0.6 minutes. In 27 cases, we used the common iliac artery and common iliac vein for vascular anastomosis. In 12 cases, the anastomosis was performed to the aorta and vena cava. Seven patients had prior augmentation cystoplasty, and the ureter was anastomosed to the pouch directly. All grafts functioned immediately, with a mean creatinine at 24 hours of 1.5 +/- 0.3 mg/dL. At last follow-up (mean 13.6 months), the mean creatinine was 0.9 mg/dL. One patient lost the graft due to severe rejection that was resistant to antithymocyte globulin. CONCLUSIONS: Laparoscopic donor nephrectomy for pediatric recipients is safe and provides quality organs with excellent function. Outcome is comparable to those after open donor nephrectomy.

Screening for coronary heart disease in elderly men based on current and past cholesterol levels.

Efficient use of cholesterol measurements to screen for coronary heart disease in the elderly is not well defined. The purpose of this report is to examine such screening based on national guidelines in a sample of older men. Since relations between cholesterol and coronary heart disease are better established in those who are younger, screening in the elderly will also consider levels of cholesterol that existed earlier in life. Data are from a prospective study of 1,170 men enrolled in the Honolulu Heart Program who were followed over a 12-year period for coronary heart disease. Follow-up began from 1980 to 1982, when cholesterol levels were determined in men who were aged 61 to 81 years. Past cholesterol levels were measured 10 years earlier (1970-1972). During the course of follow-up, coronary heart disease developed in 117 of the men. Risk of disease rose significantly (P = 0.003) with increases in past cholesterol levels (1970-1972) but not with more recent levels (1980-1982). For men with current cholesterol levels that were desirable (<5.2 mmol/L [200 mg/dl], as defined by guidelines from the National Cholesterol Education Program), disease incidence continued to rise with increasing past cholesterol levels (P<0.001). Accounting for high-density lipoprotein cholesterol and other screening factors did little to alter these findings. We conclude that desirable cholesterol levels in the elderly may not be a marker of a healthy risk profile if past cholesterol levels were high. Screening for coronary heart disease in the elderly could be improved by considering past cholesterol levels, rather than just a single measurement in later life.

Effects of walking on coronary heart disease in elderly men: the Honolulu Heart Program.

BACKGROUND: Effects of walking on the risk of coronary heart disease morbidity and mortality have not been identified in the elderly. The purpose of this study was to determine whether walking is associated with a reduced risk of coronary heart disease in a sample of elderly men. METHODS AND RESULTS: For this study, distance walked (mile/d) was examined at a baseline examination that occurred from 1991 to 1993 in the Honolulu Heart Program. Incident coronary heart disease from all causes was observed over a 2- to 4-year follow-up period. Subjects followed up were 2678 physically capable elderly men aged 71 to 93 years. During the course of follow-up, 109 men developed coronary heart disease. Men who walked <0.25 mile/d had a 2-fold increased risk of coronary heart disease versus those who walked >1. 5 mile/d (5.1% versus 2.5%; P<0.01). Men who walked 0.25 to 1.5 mile/d were also at a significantly higher risk of coronary heart disease than men who walked longer distances (4.5% versus 2.5%; P<0. 05). Adjustment for age and other risk factors failed to alter these findings. CONCLUSIONS: Findings from the Honolulu Heart Program, which targeted physically capable elderly men, suggest that the risk of coronary heart disease is reduced with increases in distance walked. Combined with evidence that suggests that an active lifestyle reduces the risk of cardiovascular disease in younger and more diverse groups, this suggests that important health benefits could be derived by encouraging the elderly to walk.

Changes in total and high-density lipoprotein cholesterol over 10- and 20-year periods (the Honolulu Heart Program).

Limited data are available on patterns of change in lipids and lipoproteins as persons age. The purpose of this report is to describe the 10-year change in total and high-density lipoprotein (HDL) cholesterol according to suspected determinants in 898 Japanese-American men enrolled in the Honolulu Heart Program. Data are based on examinations that occurred from 1970 to 1972 and at repeat examinations received 10 and 20 years later. At the last examination, men were aged 71 to 93 years. Mean reductions in total cholesterol in the second 10 years of follow-up (24 mg/dl) were more than double the reductions observed in the first 10 years (9 mg/dl). Levels of total cholesterol declined and levels of HDL cholesterol increased regardless of beginning levels of systolic blood pressure, body mass index, physical activity, cigarette smoking status, or the use of treatment for hypertension or elevated total cholesterol. Men with prevalent coronary heart disease experienced greater reductions (p <0.05) in total cholesterol during the second 10 years of follow-up (32 mg/dl) versus men without coronary heart disease (22 mg/dl). Adjustment for baseline covariates failed to alter these findings appreciably. We conclude that alterations in total and HDL cholesterol with advancing age may be expected to occur regardless of risk factor status, disease prevalence, or pharmacologic intervention. In the presence of such effects, evaluation of treatment programs to alter levels of total and HDL cholesterol in older persons should consider the possibility that even in the absence of intervention, changes could also occur due to aging alone.

Coffee consumption in hypertensive men in older middle-age and the risk of stroke: the Honolulu Heart Program.

OBJECTIVE: To examine the association between coffee consumption and the development of stroke in men at high risk for cardiovascular disease. METHODS: Coffee intake was observed from 1965 to 1968 in a cohort of men enrolled in the Honolulu Heart Program with follow-up for incident stroke over a 25-year period. Subjects were 499 hypertensive men (having systolic or diastolic blood pressures at or above 140 and 90 mm Hg, respectively) in older middle-age (55 to 68 years) when follow-up began. Past and current cigarette smokers were excluded from follow-up. RESULTS: In the course of follow-up, 76 men developed a stroke. After age-adjustment, risk of thromboembolic stroke increased significantly with increases in coffee consumption (P = 0.002). No relationships were observed with hemorrhagic stroke. When adjusted for other factors, the risk of thromboembolic stroke was more than doubled for men who consumed three cups of coffee per day as compared to nondrinkers of coffee (RR = 2.1; 95% CI = 1.2-3.7). CONCLUSIONS: Although in need of further confirmation, consumption of coffee appears to be positively associated with an increased risk of thromboembolic stroke in hypertensive men in older middle-age. Findings suggest that it may be prudent to advise older middle-aged men with hypertension who consume large amounts of coffee to consider reducing their coffee intake.

Effects of walking on mortality among nonsmoking retired men.

BACKGROUND: The potential benefit of low-intensity activity in terms of longevity among older men has not been clearly documented. We examined the association between walking and mortality in a cohort of retired men who were nonsmokers and physically capable of participating in low-intensity activities on a daily basis. METHODS: We studied 707 nonsmoking retired men, 61 to 81 years of age, who were enrolled in the Honolulu Heart Program. The distance walked (miles per day) was recorded at a base-line examination, which took place between 1980 and 1982. Data on overall mortality (from any cause) were collected over a 12-year period of follow-up. RESULTS: During the follow-up period, there were 208 deaths. After adjustment for age, the mortality rate among the men who walked less than 1 mile (1.6 km) per day was nearly twice that among those who walked more than 2 miles (3.2 km) per day (40.5 percent vs. 23.8 percent, P=0.001). The cumulative incidence of death after 12 years for the most active walkers was reached in less than 7 years among the men who were least active. The distance walked remained inversely related to mortality after adjustment for overall measures of activity and other risk factors (P=0.01). CONCLUSIONS: Our findings in older physically capable men indicate that regular walking is associated with a lower overall mortality rate. Encouraging elderly people to walk may benefit their health.

Cross-sectional and longitudinal changes in total and high-density-lipoprotein cholesterol levels over a 20-year period in elderly men: the Honolulu Heart Program.

PURPOSE: The purpose of this report is to describe levels of total cholesterol and high-density-lipoprotein cholesterol (HDL-C) in a group of elderly men and to compare these levels to those that were observed 20 years earlier. METHODS: From 1965-1968, the Honolulu Heart Program began following 8006 men of Japanese ancestry living on the island of Oahu, Hawaii, in a prospective study of coronary heart disease and stroke. This report presents data for 971 men who participated in a separate fasting study of lipids and lipoproteins that first occurred from 1970-1972 and in those who received repeat examinations 10 and 20 years later. Men were aged 71-93 years at the last examination. RESULTS: Over the 20-year period, total cholesterol declined by 1.6-1.8 mg/dL per year (P < 0.001), from average baseline values of 219-222 mg/dL. Levels of HDL-C rose 0.2-0.3 mg/dL per year (P < 0.001), from average baseline values of 44-46 mg/dL. After adjustment for baseline cholesterol levels, men with prevalent coronary heart disease at the end of the 20-year follow-up experienced significantly greater reductions in total cholesterol levels than men without disease (P < 0.001). Men who developed coronary heart disease within the first 10 years of follow-up had the greatest yearly decline in total cholesterol (1.9 mg/dL), followed by men who developed heart disease later (1.8 mg/dL) and men who remained disease free (1.5 mg/dL). Differences between men with recent and earlier disease were not statistically significant, although men without coronary disease experienced a significantly smaller decrease in total cholesterol than either of these groups (P < 0.05). CONCLUSIONS: Changes in total cholesterol and HDL-C levels with advancing age may be part of a natural aging process. Some changes, however, such as large reductions in total cholesterol, may signal occult disease or declines in overall health. Selective survival may contribute to these findings since improvements in lipid and lipoprotein levels that are beneficial in younger ages were common in this long-lived cohort of men.

Endocrine abnormalities in patients with amyloidosis.

This study was conducted to examine systematically the endocrine function in 10 patients diagnosed to have renal amyloidosis. Basal and dynamic endocrine tests (GnRh, TRH, and ACTH stimulation tests as well as modified glucose tolerance test) were performed in all patients. Eight patients had advanced renal disease (creatinine clearance < 10 mL/min). Hypofunction of the thyroid gland was present in 5 out of those 8 patients, and thyroid biopsy confirmed amyloid deposition. Of those 5 patients, pituitary and adrenal defects were present in 2. In conclusion, endocrine abnormalities were evident at a later stage of renal amyloidosis. Hypothyroidism was present in all patients who manifested endocrine defects and should be considered as an early sign of endocrine involvement with the disease.

Computer-aided interpretation of ECG signals using polynomial regression methods.

Using polynomial regression methods this paper introduces a new approach to computer-aided interpretation of the ECG. An algorithm is presented for the identification and separation of ECG segments. The method of interpretation is efficient and its results are consistent and reliable.

Prognostic significance of mRNA-encoding estrogen receptor and epithelial growth factor receptor in breast carcinoma progression into lymph nodes: 1. Estrogen receptor encoding mRNA.

The main objective of this study was to differentiate between lymph nodes infiltrated by estrogen receptor-positive (ER+) and estrogen receptor-negative (ER-) breast carcinoma. Lymph nodes were obtained from 40 postmenopausal cancer patients, 10 from each disease stage. Six patients from each group had estrogen receptor-positive (BCaER+) and four estrogen receptor-negative (BCaER-) tumors. Both tumor-containing (T) and uninvolved (N) lymph nodes from the same patient were examined by the following parameters: magnitude of lymph node nucleic acid hybridization with cDNA probes from breast cancer MCF-7ER+ and MCF-7ER- cells; and binding capacity of 3H-estradiol, 125I-EGF, and 125I-PDGF binding and protein kinase C activities of the lymph nodes. Concomitant with the appearance of transformed cells, several events occur: Tumor cells induce stimulation of mononuclear cells and macrophages and evoke T- and B-cell proliferation, leading to the synthesis of tumor cell membrane-associated antibodies. In estrogen receptor-positive (ER+) breast carcinoma, estrogens and host hormonal modulatory mechanisms stimulate production and release of epithelial growth (EGF) and platelet-derived growth factors (PDGF). These factors are characterized by protein kinase C activities. There is infiltration of tumor cells into the lymph node and infiltration of leukocytes into the tumor site. In the lymph node, tumor progression depends on tumor cell proliferation rate and metastatic aggressiveness. The experiments described in this study document the changes that occur in lymph nodes, with differences between nodes infiltrated with BCaER+ and BCaER- breast carcinomas. Hybridization of 32P-cDNA from MCF-7ER+ cells with cellular RNA from BCaER+ involved (T) lymph nodes is greater than with cellular RNA from uninvolved (N) lymph nodes. The magnitude of hybridization correlated (P less than 0.005) with the disease stage.

Peripheral blood lymphocytes from patients with cancer lack interleukin-2 receptors.

When tumor cells develop in healthy adults, they activate the cellular immune system--natural killer (NK) cells, antigen-specific cytotoxic lymphocytes (CTL), and the synthesis of antigen specific cytotoxic antibodies. These are aimed at killing the intruding cells. However, in cancer patients the tumor continues to grow. As tumor cells proliferate, they were shown to release factors that mediate the inactivation of the host immune defense systems. The study documented in this article examined peripheral blood lymphocytes, mononuclear cells (MNC), NK cells, T-helper cells (THC). This study confirmed the interaction of the released inhibitor factors with these mononuclear cells. NULL cells from healthy adults responding to interleukin-2 (IL-2) and NILL cells from patients with metastatic breast carcinoma nonresponsive to IL-2 were also isolated by the standard antibodies-pinning technique. The cells were obtained from age-matched subjects: ten healthy adults; ten patients each from Stage I, II, III, and IV metastatic breast carcinoma (BCa-I, BCa-II, BCa-III, and BCa-IV or MBCa); and ten patients with benign breast disease (BBD). The responsiveness of these THC, PBMNC, NK, NULL, and NILL cells in vitro to graded levels of phytohemagglutinin (PHA), Concanavalin A (Con A), and recombinant interleukin-2 (rIL-2) was examined. Responsiveness was monitored by 3H-thymidine (3H-TdR) uptake, production and release of IL-2, interleukin-2 receptor (IL-2R), and cytotoxic activities against K-562 cells and breast carcinoma short-term cell lines. A lack of functional IL-2R in peripheral blood lymphocytes from patients with metastatic breast carcinoma was confirmed by nonsignificant anti-Tac antibody binding. An elevation in the expression of cell surface antigen GP-120 has been observed to be associated with the activation in vitro of T-cells from healthy adults and from patients with benign breast disease, but not of T-cells from patients with breast carcinoma. Biochemical studies of the GP-120 using high performance liquid chromatography combined with nitrocellulose blotting confirmed that the glycoprotein was resistant to trypsin and chymotrypsin, but susceptible to pronase. It contained sialic acid and lactosaminoglycan as O-linked sugars. It could be labeled with pariodate/NaB(3H4) and is recognized by MAbT-305 monoclonal antibodies. It contained sialic acid linked (2---3) to galactose.(ABSTRACT TRUNCATED AT 400 WORDS)

A mechanism by which human breast carcinoma cells escape the host immune system.

The experiments described in this study examined cell membrane oligosaccharides, malignancy-related cell phenotypes and tumor cell susceptibility to the killing effect of human cytotoxic cells. Short term breast carcinoma (BCa) cell lines were prepared from biopsies obtained from patients at each of the pathological Stages I, II, III and from patients with disseminated liver metastasis. Five patients at each stage donated the tissue. To obtain large enough quantities, the cells were cultured as monolayers for a brief period, then transferred to roller bottles using serum-free hormone defined medium. Natural killer (NK) cells, lymphokine (IL-2)-activated killer (LAK), tumor-infiltrating lymphocytes (TIL) and peripheral cytotoxic lymphocytes (CTL) from patients with BCa at PS I were used as the effector cells. Susceptibility of the tumor cells to the killing effects of the effector cells was monitored by the well established 4 h 51Cr-release assay technique. Growth factor expression, oncogenicity in athymic female mice and colonigenicity in soft agar were used as parameters to monitor breast carcinoma cell malignancy phenotypes. The cell membrane oligosaccharides were determined from the carbohydrate elution profiles from BioGel P-6 columns. The results indicate a correlation between progression of malignancy from PS I to the metastatic stage PS IV, and the magnitude of malignancy phenotypes, resistance to the host killer cells and oligosaccharide profile shift to a higher molecular size with increased sialylation and fucosylation of the carbohydrate moieties.