Islet cell metabolism is reflected by the MTT (tetrazolium) colorimetric assay.

Insulin secretion depends critically on glucose metabolism. We investigated whether a rapid viability test could be established for assessing glucose metabolism in insulin secreting cells. The MTT (C,N-diphenyl-N'-4,5-dimethyl thiazol-2-yl tetrazolium bromide) colorimetric assay (reduction of tetrazolium salt to formazan) was applied to rat islets and rat insulinoma cell lines. It was found that the rate of formazan production correlated with glucose oxidation and glucose utilization at glucose concentrations which also stimulated insulin secretion. In differentiated insulinoma INS-1 cells, salt reduction paralleled the insulin release at glucose concentrations of up to 8.3 mmol/l. The glucose-induced formazan production in INS-1 cells and islets was abolished by exposure to the Beta-cell cytotoxic agents, streptozotocin or alloxan. The MTT assay thus provides a convenient tool for the rapid assessment of Beta-cell metabolism and viability.


Dehydroepiandrosterone: the "missing link" between hyperinsulinemia and atherosclerosis?

A well-established epidemiologic association exists between hyperinsulinemia and macrovascular disease. However, the mechanism or mechanisms by which hyperinsulinemia promotes atherogenesis is unknown. Recent evidence indicates that the adrenal steroid dehydroepiandrosterone (DHEA) exerts multiple antiatherogenic effects and also suggests that hyperinsulinemia may reduce serum DHEA and DHEA-sulfate levels by decreasing production and enhancing metabolic clearance. We advance the hypothesis that hyperinsulinemia promotes macrovascular disease in part by reducing serum DHEA and DHEA-sulfate levels and illustrate how this may be the case in two clinical conditions characterized by hyperinsulinemic insulin resistance: aging and obesity.


Isolation and partial characterization of a compound with siderophore activity from Vibrio parahaemolyticus.

A compound with siderophore activity was purified by successive column and thin layer chromatographic procedures from Dowex 1 x 8 extracts of culture supernatants of Vibrio parahaemolyticus AQ 3354. The strain synthesized the compound in culture media containing less than 2 microM added FeCl3. Hydrolysis of the compound yielded alanine, ethanolamine, citric acid and 2-ketoglutaric acid. The 1H-NMR spectrum exhibited the presence of a residue from each of these components in the intact molecule. The fast-atom bombardment mass spectrum of the methyl ester derivative indicated a prominent ion at m/z 477, probably corresponding to [M + 1] ion. Other strains of V. parahaemolyticus were also found to produce this compound when grown in an iron-limited medium.


Rhizoferrin: a complexone type siderophore of the Mucorales and entomophthorales (Zygomycetes).

The present investigation presents evidence that rhizoferrin, a novel polycarboxylate or complexone-type siderophore, originally isolated from Rhizopus microsporus, represents the common siderophore within the Zygomycetes. Thus, rhizoferrin could be detected by HPLC analysis in various families of the Mucorales, e.g., Rhizopus microsporus var. rhizopodiformis, Mucor mucedo and Phycomyces nitens (Mucoraceae), Chaetostylum fresenii and Cokeromyces recurvatus (Thamnidiaceae), Cunninghamella elegans and Mycotypha africana (Choanephoraceae) and Mortierella vinacea (Mortierellaceae) and in Basidiobolus microsporus (Entomophthorales). The function of rhizoferrin as a siderophore in the fungus R. microsporus var. rhizopodiformis was demonstrated by time- and concentration-dependent uptake of [55Fe]-labelled rhizoferrin, yielding saturation kinetics with values of Km = 8 microM and V(max) = 1.2 nmol min-1 (mg dry wt)-1.


Specificity and sensitivity of hexosaminidase assays and DNA analysis for the detection of Tay-Sachs disease gene carriers among Ashkenazic Jews.

Tay-Sachs disease (TSD), a neurodegenerative disorder resulting from a deficiency of the lysosomal enzyme hexosaminidase A (HexA), clusters in Ashkenazic Jews. Population-based screening programs to detect carriers of TSD genes by means of HexA assays have been active since the 1970s. The recent characterization of 3 mutations in the HEXA gene (in exon 7, exon 11, and intron 12), which account for over 90% of HEXA mutations in Ashkenazim, appeared to offer better options for screening and diagnosis. The relative frequencies of the three mutations in Montreal are similar to those reported in four other North American populations. We compared enzyme and DNA analyses to determine specificity and sensitivity of each test when the other was used as the confirmatory procedure. Neither procedure has a sensitivity of 1.0. Maximum sensitivity and specificity were achieved by using both tests together. The findings here are likely to apply to most cases where the variant screened enzyme phenotype can result from more than one mutation.


Coagulopathy in disseminated intravascular coagulation due to abdominal sepsis: determination of prothrombin fragment 1 + 2 and other markers.

To estimate the degree of coagulopathy in abdominal sepsis, we measured the plasma levels of prothrombin fragment 1 + 2 (F1 + 2), thrombin-antithrombin III complex (TAT) and plasmin-alpha 2-plasmin inhibitor complex (PIC) by the enzyme-linked immunosorbent assay in 38 patients with disseminated intravascular coagulation (DIC). In 20 patients with DIC due to abdominal sepsis, plasma levels of F1 + 2, TAT and PIC were 2.6 nmol/l, 27.9 micrograms/l and 1.5 micrograms/ml, respectively, with a mean antithrombin III (AT III) activity of 41.7%. F1 + 2, TAT, PIC and AT III levels were 4.7 nmol/l, 75.8 micrograms/l, 8.8 micrograms/ml and 70.9% in 18 patients with DIC as the result of malignancy. Though AT III levels in DIC due to sepsis were lower than those in DIC due to malignancy, the levels of F1 + 2, TAT and PIC in the former were not significantly more increased than those in the latter. The plasma levels of F1 + 2 were positively correlated with TAT and PIC in DIC patients with malignancy; however, there was no correlation between F1 + 2 and TAT or PIC in DIC patients with sepsis. In addition, the levels of serum albumin in the two groups were similar. These results suggest that activation of coagulation and fibrinolytic systems may not be so prominent in cases of DIC due to abdominal sepsis, compared to related events in DIC due to malignancy. It is also suggested that the depletion of AT III in cases of sepsis is not only caused by a consumption related to intravascular coagulation or to an alternate distribution of protein.(ABSTRACT TRUNCATED AT 250 WORDS)


[Severe disability. Part 2: The difficult comparison with same age probands].

With their texts on Schwerpflegebedürftigkeit, the legislators have set the interpreters of legal texts a difficult task. In practice, attempts by the "users" of such texts to find solutions inexorably lead to aporias. Responsible for decision-taking is the health insurance carrier, with the physician exclusively acting as a counsellor on the medical aspects. The advised comparisons with persons of identical age with the intention of establishing Schwerpflegebedürftigkeit is not required by the legislator, and it is not easy to see what sense it is supposed to make. The corresponding guidelines drawn up by the leading associations are not unequivocal and are therefore open to misunderstanding.


Nutritional alterations in persons with HIV infection.

Potential relationships among nutritional status, immune function and quality of life were examined in a convenience sample of 40 outpatient homosexual and bisexual males stratified into five categories, using modified Walter Reed Staging Criteria. Nutritional status was assessed by measuring height, weight, triceps skinfold thickness, arm circumference, nutrient intake and serum albumin. Immune status was evaluated by determining T-helper cell numbers and percentages. The Quality of Life test was used to obtain information about life quality. Nutritional assessment failed to show significant differences among groups with the exception that serum albumin levels were reduced in persons with AIDS. The significance of change in serum albumin in regard to nutritional status is unclear, since serum albumin is affected by a number of non-nutritional factors, such as hydration status and liver function. The study also revealed a significant decline in T-helper percentages, but not absolute T-helper cell numbers as a function of disease stage. There were no statistically significant differences between the quality of life scores with respect to each grouping. These data suggest that asymptomatic patients as well as those with ARC or stable AIDS are able to maintain body weight and composition.


In vitro endothelialization of carbon-coated Dacron vascular grafts.

Physical vapour deposition is used to coat vascular prostheses with pyrolytic carbon. This coating may facilitate the development of an endothelial monolayer in grafts implanted in laboratory animals. This in vitro study compared the adherence and growth of cultured porcine aortic endothelial cells (EC) seeded in vitro on carbon-coated Dacron, to uncoated prostheses. The cells were incubated at 5 x 10(4) cells/cm2. Progress was monitored at different times (TO + 2 hours, D 1, D 4, D 8) by microscopic observation, when cell counts were made; and by scanning electronic microscopy (SEM) to evaluate morphological EC-graft interactions. Cell adherence was independent of the carbon coating but cellular growth occurred only on carbon-coated Dacron. The SEM observations showed both the shape of the adherent cells, which were rounded on uncoated Dacron and extensively spread on carbon-coated prostheses, and the morphology of the carbon coating.


The human squamous cervical carcinoma cell line, HOG-1, is responsive to steroid hormones.

Growth of the human squamous cervical carcinoma cell line, HOG-I, was stimulated in response to oestradiol in serum-containing and chemically defined medium. The oestradiol-stimulated growth could be inhibited by 4-OH tamoxifen, progesterone and medroxyprogesterone acetate; the last 2 compounds also inhibited basal cell growth in serum-containing and chemically defined media. The data are consistent with the sensitivity of human squamous cervical cancer to sex-steroid hormones and suggest that endocrine therapies may be of benefit in this disease.


Deoxyspergualin, a novel immunosuppressant, markedly inhibits human mixed lymphocyte reaction and cytotoxic T-lymphocyte activity in vitro.

Deoxyspergualin (DSG) has demonstrated potent immunosuppressive activities in vivo. However, because of its lability in culture medium, the mechanism of activity has not yet been identified in vitro. In this study, a more stable analogue, deoxymethylspergualin (MeDSG), was used to investigate the in vitro immunosuppressive activity of DSG. MeDSG suppressed both human mixed lymphocyte reaction (MLR) and cytotoxic T-lymphocyte (CTL) activity at doses greater than 0.1 micrograms/ml in vitro. In kinetics studies, MeDSG was found to suppress a MLR when added on day 3 of a 7 day MLR incubation but cyclosporin A (CYA) suppressed a MLR only when added during the initial stage of a MLR (i.e. on day 1). In studies of cell surface phenotype in the MLR, MeDSG treatment decreased the numbers of CD8+ lymphocytes but those of CD4+ lymphocytes were not affected. In addition, MeDSG had no significant effect on interleukin-2 (IL-2) receptor expression or IL-2 production. These results suggest that MeDSG suppresses the T-cells which are proliferating competent cells such as cytotoxic T-cells (CD8+), but has a different mode of immunosuppressive action compared with CYA.


Effects of a diet regimen on pituitary and steroid hormones in male ice hockey players.

Serum concentrations of androgens, cortisol, androgen binding proteins, pituitary hormones, together with anthropometric variables and sports performance were studied in two different elite male ice hockey teams. One of the teams (DIF, n = 22) participated in a special dietary program including reduction in fat from approximately 40 per cent of total energy intake (E%) to less than 30 E% and an increase in carbohydrate intake from 45 E% to about 55 E%, while the other (SSK, n = 21) served as a control group and had no special dietary program. The study covered a 7-month period. Basal values of serum testosterone, sex hormone binding globulin (SHBG), non-SHBG-bound testosterone (NST), cortisol, dehydroepiandrosterone sulfate (DHAS) and LH did not differ between the two teams. Serum concentrations of testosterone, SHBG, NST and cortisol increased significantly during the study period in the DIF group and were, with the exception of SHBG, significantly higher than in the SSK group at the end of the study (33.0 vs 26.8 nmol/l, p less than 0.05; 22.5 vs 18.3 nmol/l, p less than 0.05; and 548 vs 464 nmol/l, p less than 0.01). The ratio between NST and cortisol which was used as an index of anabolic/catabolic steroid balance did not change in either group during the study. A significant decrease in the serum concentrations of LH during the observation period was found in the SSK group. The endocrine differences between the teams may be explained by a relative negative energy balance in DIF, together with a reduced fat and increased carbohydrate intake.(ABSTRACT TRUNCATED AT 250 WORDS)


The relationship between age and the prevalence of senile dementia: a meta-analysis of recent data.

A linear regression model derived from a meta-analysis of 13 epidemiological studies of senile dementia conducted since 1980, and employing internationally-known case-finding procedures, suggests a much lower general rate of dementia prevalence than has been previously estimated. An exponential increase with age is observed, with senile dementia prevalence diagnosed by Diagnostic and Statistical Manual (DSM-III) criteria doubling every 6 years and senile dementia of the Alzheimer's type (SDAT) every 4.2 years. Studies providing data for the oldest ages indicating a drop in the rate of increase after the age of 80 suggest that senile dementia may be age-related rather than ageing-related. Estimates derived from this model may provide a reasonably accurate means of estimating dementia prevalence in the general population. The limitations of this method for the purposes of prediction and studies of risk factors are discussed in relation to the hypothesized heterogeneity of senile dementia and possible cohort effects.


Chronic liver disease rarely follows acute hepatitis B in non-immunocompromised adults.

The risk of developing a chronic carriage state after acute hepatitis B infection in adults was evaluated. Two hundred and eighty-nine HBV-susceptible heterosexual partners of acute hepatitis B patients were used to investigate the effectiveness of post-exposure immunoprophylaxis; 75 of them received hepatitis B vaccine, 72 hepatitis B hyperimmune globulin (HBIG), 71 vaccine plus HBIG and 71 placebo. Participants were interviewed, clinically examined and serum specimens were taken at 1, 3, 6 and 9 months after their first intervention. Serum samples were tested for ALT and HBV markers (HBsAg, anti-HBc and anti-HBs) using radio immunoassays. Forty-six (15.9%) of the heterosexual partners examined were infected; the incidence of HBV infections was higher among placebo (18.3%, 13/71) and HBIG (18.1%, 13/72) recipients compared to vaccine (16.0%, 12/75) and HBIG plus vaccine (11.3%, 8/71) recipients, but the differences were not statistically significant. Infections were significantly more often subclinical after immunoprophylaxis (p = 0.03). HBsAg was detected in all eight clinical and in 13 of the 38 subclinical cases. In the remaining 25 subclinical cases HBV infections were diagnosed by the development of anti-HBc and anti-HBs during the follow-up period. Finally, all 46 cases studied cleared the HBsAg.


Tumor diagnosis by PET: potential of seven tracers examined in five experimental tumors including an artificial metastasis model.

The potential of seven tracers for the metabolic imaging of tumors by positron emission tomography was studied using five experimental tumor models. The tracers examined were 2-deoxy-2-[18F]fluoro-D-glucose ([18F]FDG), 2-deoxy-2-[18F]fluoro-D-galactose (2-[18F]FdGal) and 2-deoxy-2-[18F]fluoro-L-fucose (2-[18F]FdFuc) for investigating energy metabolism. L-[methyl-11C]Methionine ([11C]Met) and 6-[18F]fluoro-L-fucose (6-[18F]FFuc) were used for assessing protein and glycoprotein synthesis, while [3H]thymidine ([3H]Thd) and 2-deoxy-5'-[18F]fluorouridine ([18F]FdUrd) were used to investigate nucleic acid metabolism. The highest mean uptake by the five different tumors was found for [3H]Thd, followed in order by [18F]FDG, [11C]Met, 2-[18F]FdGal, [18F]FdUrd, 2-[18F]FdFuc and 6-[18F]FFuc. The tumor-to-tissue uptake ratios indicated that the nucleosides, [11C]Met and 6-[18F]FFuc were better tracers in the brain region. All the tracers except for the fucose analogs were suitable for the thoracic region, while [11C]Thd and [18F]FDG were superior in the abdominal region. In comparison with the primary tumor model of Lewis lung carcinoma (3LL), [3H]Thd uptake in the artificial metastatic 3LL model showed the maximum enhancement, followed by [18F]FDG, [11C]Met and the other tracers. The [18F]FDG uptake correlated with the [3H]Thd uptake. [18F]FdUrd, 6-[18F]FFuc and 2-[18F]FdGal could be used for distinguishing different types of tumors. The combined use of these radiotracers can possibly allow the assessment of tumor metabolism, and this indicates the viability of tumors.


[Demonstration of soluble IgG-Fc type II receptors (or s-Fc-gamma-II-R) in human whole saliva].

Twenty human salivary samples from two groups of patients (with or without dental caries) were tested for the presence of soluble forms of Fc gamma receptors type II (sFc gamma RII): sFc gamma RII were detected by immunoblotting using a radiolabelled monoclonal antibody directed against human Fc gamma RII. These soluble forms specifically interact with the Fc portion of IgG and are produced by cells of the immune system (macrophages, neutrophils, Langerhans cells). This study showed that sFc gamma RII are present in human unstimulated saliva. The statistical analysis indicated that the relative amounts of sFc gamma RII in human unstimulated saliva are variable depending on the patient. No apparent association was found between the sFc gamma RII and the oral status of the patients.


Cytoplasmic dynein participates in the centrosomal localization of the Golgi complex.

The localization of the Golgi complex depends upon the integrity of the microtubule apparatus. At interphase, the Golgi has a restricted pericentriolar localization. During mitosis, it fragments into small vesicles that are dispersed throughout the cytoplasm until telophase, when they again coalesce near the centrosome. These observations have suggested that the Golgi complex utilizes a dynein-like motor to mediate its transport from the cell periphery towards the minus ends of microtubules, located at the centrosome. We utilized semi-intact cells to study the interaction of the Golgi complex with the microtubule apparatus. We show here that Golgi complexes can enter semi-intact cells and associate stably with cytoplasmic constituents. Stable association, termed here "Golgi capture," requires ATP hydrolysis and intact microtubules, and occurs maximally at physiological temperature in the presence of added cytosolic proteins. Once translocated into the semi-intact cell cytoplasm, exogenous Golgi complexes display a distribution similar to endogenous Golgi complexes, near the microtubule-organizing center. The process of Golgi capture requires cytoplasmic tubulin, and is abolished if cytoplasmic dynein is immunodepleted from the cytosol. Cytoplasmic dynein, prepared from CHO cell cytosol, restores Golgi capture activity to reactions carried out with dynein immuno-depleted cytosol. These results indicate that cytoplasmic dynein can interact with isolated Golgi complexes, and participate in their accumulation near the centrosomes of semi-intact, recipient cells. Thus, cytoplasmic dynein appears to play a role in determining the subcellular localization of the Golgi complex.


The inner dynein arms I2 interact with a "dynein regulatory complex" in Chlamydomonas flagella.

We provide indirect evidence that six axonemal proteins here referred to as "dynein regulatory complex" (drc) are located in close proximity with the inner dynein arms I2 and I3. Subsets of drc subunits are missing from five second-site suppressors, pf2, pf3, suppf3, suppf4, and suppf5, that restore flagellar motility but not radial spoke structure of radial spoke mutants. The absence of drc components is correlated with a deficiency of all four heavy chains of inner arms I2 and I3 from axonemes of suppressors pf2, pf3, suppf3, and suppf5. Similarly, inner arm subunits actin, p28, and caltractin/centrin, or subsets of them, are deficient in pf2, pf3, and suppf5. Recombinant strains carrying one of the mutations pf2, pf3, or suppf5 and the inner arm mutation ida4 are more defective for I2 inner arm heavy chains than the parent strains. This evidence indicates that at least one subunit of the drc affects the assembly of and interacts with the inner arms I2.


Phorbol ester-induced promyelocytic leukemia cell adhesion to marrow stromal cells involves fibronectin specific alpha 5 beta 1 integrin receptors.

The human promyelocytic cell line NB4 exhibited a weak adhesion capacity for bone marrow-derived stromal cells and their extracellular matrices (5-15% of adherent cells). Adhesion was enhanced by pulse-treatment of cells with phorbolester (PMA 10(-7) M). Adhesion was induced within minutes, was fibronectin-specific, and affected up to 100% of the treated cells. This biological response to PMA resulted from the activation of protein kinase C (PKC), since PKC inhibitors (staurosporine, sphingosine, CGP 41251, and calphostin C) prevented the phenomenon. Phenotypical analysis of integrin receptor expression (particularly FN receptors VLA-4 and VLA-5) at the membrane of untreated or PMA-treated cells revealed that PMA induced no significant modification of the level of expression of these receptors. However, inhibition studies carried out with anti-VLA monoclonal antibodies demonstrated that the FN-specific adhesion triggered by PKC involved the alpha 5 beta 1 FN-specific receptors (VLA-5). We showed that the binding of NB4 cells to fibronectin was RGD-dependent. PMA-induced adhesion was not correlated to phosphorylation of the VLA-5 receptor. These findings may partially explain the malignant behaviour of these cells: The loss of their capacity to adhere to stromal cells may arrest differentiation and explain the large number of leukemic cells in the circulation.


Cytoplasmic accumulation of cyclin B1 in human cells: association with a detergent-resistant compartment and with the centrosome.

Mitotic cyclins are thought to function as key regulatory subunits of the universal M-phase-promoting p34cdc2 protein kinase. Previous immunolocalization studies have demonstrated that a fraction of p34cdc2 undergoes cell cycle-dependent accumulation at the centrosome during G2/M. In order to identify the mitotic cyclins with which this p34cdc2 fraction interacts, we carefully examined the subcellular distribution of both cyclin A and B1 in HeLa cells. We show here that part of cyclin B1 is associated with duplicating centrosomes throughout its accumulation in the cytoplasm and up to metaphase. In contrast cyclin A does not exhibit centrosomal association except at the onset of mitosis, from preprophase up to metaphase. We also present cytological and biochemical evidence that cyclin B1 is preferentially accumulated as a detergent-insoluble protein independently of the state of microtubule assembly and under experimental conditions where most of p34cdc2 is soluble. Interestingly, the electrophoretic pattern of the minor insoluble p34cdc2 fraction was previously shown to be particularly enriched in slow-migrating and presumably hyperphosphorylated isoforms, known to interact specifically with cyclin B1 during interphase. From these results we propose that the interaction of cyclin B1 with the centrosomes and with the cytoplasmic structures is a constitutive feature of the mechanism whereby a fraction of p34cdc2 is recruited and subsequently targeted to the cyclin B-dependent activation pathway.


Distribution of cytoplasmic and axonemal dyneins in rat tissues.

Microtubule-associated protein 1C (MAP 1C) is now defined as brain cytoplasmic dynein. Recent studies have suggested that cytoplasmic dynein is a motor protein responsible for the intracellular microtubule-based motility in neuronal and non-neuronal cells. We have prepared an antibody against bovine brain MAP 1C and have examined the localizations of cytoplasmic dynein in rat tissues. Immunoblots of extracts from the tissues showed that the dynein was present in brain, testis, liver, kidney and lung. Immunohistochemical experiments have demonstrated that dynein is localized in Purkinje cells of cerebellum and axons of central and peripheral nervous systems. In non-neuronal tissues, the antibody staining was intense in many types of cells, such as hepatocytes, epithelia of renal convoluted tubules, secretory cells of adrenal medulla and spermatids. Glomeruli of kidney, bronchial epithelia and type II pneumocytes of lung, pancreatic islets and acini, adrenal cortex and Sertoli cells were moderately positive upon exposure to the cytoplasmic dynein antibody. On the other hand, the localization of axonemal dynein was examined using antibodies against flagellar dynein of sea urchin spermatozoa. Anti-axonemal dynein labeled cilia and flagella in rat tissues whereas anti-MAP 1C did not stain axonemes. We also tested for immunological cross-reactivity between cytoplasmic and axonemal dyneins to probe for molecular similarities. Anti-axonemal dynein reacted with MAP 1C weakly. These results have confirmed that cytoplasmic dyneins are distributed widely among rat organs, not only in neuronal but also in non-neuronal tissues. There is no similarity in the localization of cytoplasmic and axonemal dyneins but there is some similarity in molecular antigenicity.


Implications of ultrasonically diagnosed polycystic ovaries. II. Studies of dynamic and pulsatile hormonal patterns.

Studies of 6-h hormone pulse patterns distinguished patients with polycystic ovarian disease (PCOD) from those with hyperprolactinaemia or hypothyroidism associated with ultrasonically diagnosed polycystic ovaries (PCO). No specific derangement in the gonadotrophin pulse pattern was responsible for these changes, as shown in patients with and without PCO in the latter two groups. These changes may reflect an abnormal ovarian response to normal or abnormal gonadotrophic drive. Out of 26 patients with PCO and elevated dehydroepiandrosterone sulphate (DHEA-S) levels, only three patients (11.5%) proved to have adrenal 21-hydroxylase deficiency. Ultrasonic visualization of polycystic ovaries must be supplemented with an endocrine biochemical assessment. Moreover, mild elevation of DHEA-S, without a concurrently high 17 alpha-hydroxyprogesterone level was not diagnostic of adrenal hyperplasia.


Do analogues of gonadotrophin releasing hormone influence follicular fluid steroid levels, oocyte maturity and fertilization rates?

One-hundred-and-twelve samples of follicular fluid from 32 patients undergoing in-vitro fertilization and embryo transfer were analysed in this study. The follicular fluids were analysed for any relationships between oestradiol, progesterone and 17 alpha-hydroxyprogesterone levels, the progesterone/oestradiol and 17 alpha-hydroxyprogesterone/oestradiol ratios and oocyte maturity and fertilization rates. In Group A, consisting of women who used analogues of gonadotrophin-releasing hormone during ovarian stimulation with human menopausal gonadotrophin, the progesterone/oestradiol ratio rose in parallel with the fertilization rate (P less than 0.05). Group B comprised patients treated with human menopausal gonadotrophin alone. No significant relationship was found between the other parameters, oocyte maturation and fertilization rates in either group.


Fertility following laparoscopic management of benign adnexal cysts.

Fertility following laparoscopic treatment of benign adnexal cysts without ovarian suture was studied retrospectively. Patients with endometriomas or who were previously infertile were excluded. Thirty-eight patients treated conservatively were included, 10 after partial resection of functional cysts, 23 after an ovarian cystectomy and six after treatment of a paraovarian cyst. One patient had two cysts. The overall intrauterine pregnancy rate was 92%; one patient had an ectopic pregnancy (2.6%). From these results, we conclude that fertility following laparoscopic treatment of adnexal cysts appears to be normal. Technical guidelines to improve laparoscopic cystectomy are proposed.


Cell retargeting by bispecific monoclonal antibodies. Evidence of bypass of intratumor susceptibility to cell lysis in human melanoma.

Intratumor heterogeneity for susceptibility to cytotoxic T lymphocytes (CTL)-mediated lysis represents a major obstacle to cancer adoptive immunotherapy. To overcome the heterogeneity observed in terms of susceptibility of target cells to cell-mediated lysis, in this study we used two purified bispecific monoclonal antibodies (bsmAbs) that recognize molecules expressed by cytotoxic effector cells (CD3 and IgG Fc receptorial molecules), as well as one high molecular weight melanoma-associated antigen (HMW-MAA). The ability of these reagents to enhance or induce a relevant in vitro cytotoxic activity by a CTL clone (CTL 49) isolated from PBL of a melanoma patient was tested on a large panel of autologous and allogeneic melanoma cell lines and clones. Functional studies revealed that the CTL 49 clone lysed all the HMW-MAA+ tumor lines in the presence of bsmAbs and that these reagents affected the target lysis in a cooperative fashion. The effectiveness of bsmAbs in overcoming the heterogeneous susceptibility of human melanoma cells to cell-mediated lysis may find practical implications in cancer adoptive immunotherapy.


Transfection of a glycosylated phosphatidylinositol-anchored folate-binding protein complementary DNA provides cells with the ability to survive in low folate medium.

KB cells express a folate-binding protein that is anchored to the plasma membrane by a glycosylated phosphatidylinositol (GPI) tail and these cells can grow in medium containing a very low folate concentration (1 nM). In contrast, mouse 3T3 cells do not express a membrane-associated folate-binding protein and cannot grow under similar low folate conditions. In these studies, 3T3 cells were transfected with a vector containing the cDNA that codes for the KB cell folate-binding protein. In contrast to the wild-type 3T3 cells, the transfected 3T3 cells express a level of folate-binding protein similar to KB cells, 1 and 1.4 ng/micrograms protein, respectively. The capacity for binding [3H] folate to the surface of transfected 3T3 cells cultured in folate-deficient medium is 7.7 pmol/10(6) cells, and this is approximately 50% of the surface binding capacity of KG cells under similar culture conditions. Moreover, after treatment of the transfected 3T3 cells with phospholipase C specific for phosphatidylinositol, the binding of [3H] folate to the surface of these cells is reduced by 90%, indicating that, like the KB cells, the folate-binding protein is anchored to the plasma membrane by a GPI tail. Although the doubling time of wild-type 3T3 cells markedly increases after 13 d of culture in folate-deficient medium, the doubling time of both the transfected 3T3 cells and KB cells do not change. The results of these experiments indicate that the GPI-anchored folate-binding protein provides a mechanism to maintain a level of folate that permits the folate-dependent metabolic functions necessary for cell survival under low folate conditions.


Live Borrelia burgdorferi preferentially activate interleukin-1 beta gene expression and protein synthesis over the interleukin-1 receptor antagonist.

Lyme arthritis is one of the few forms of chronic arthritis in which the cause is known with certainty. Because cytokines are thought to contribute to the pathogenesis of chronic arthritis, we investigated the effect of the Lyme disease spirochete, Borrelia burgdorferi, on the gene expression and synthesis of IL-1 beta and the IL-1 receptor antagonist (IL-1ra) in human peripheral blood mononuclear cells. Live B. burgdorferi induced fivefold more IL-1 beta than IL-1 alpha and sevenfold more IL-1 beta than IL-1ra; LPS or sonicated B. burgdorferi induced similar amounts of all three cytokines. This preferential induction of IL-1 beta was most dramatic in response to a low passage, virulent preparation of B. burgdorferi vs. three high passage avirulent strains. No difference in induction of IL-1ra was seen between these strains. The marked induction of IL-1 beta was partially diminished by heat-treatment and abrogated by sonication; IL-1ra was not affected. This suggested that a membrane component(s) accounted for the preferential induction of IL-1 beta. However, recombinant outer surface protein beta induced little IL-1 beta. By 4 h after stimulation, B. burgdorferi induced sixfold more IL-1 beta protein than LPS. In contrast to LPS-induced IL-1 beta mRNA which reached maximal accumulation after 3 h, B. burgdorferi-induced IL-1 beta mRNA showed biphasic elevations at 3 and 18 h. B. burgdorferi-induced IL-1ra mRNA peaked at 12 h, whereas LPS-induced IL-1ra mRNA peaked at 9 h. IL-1 beta synthesis increased in response to increasing numbers of spirochetes, whereas IL-1ra synthesis did not. The preferential induction by B. burgdorferi of IL-1 beta over IL-1ra is an example of excess agonist over antagonist synthesis induced by a microbial pathogen, and may contribute to the destructive lesion of Lyme arthritis.


Percutaneous arterial gene transfer in a rabbit model. Efficiency in normal and balloon-dilated atherosclerotic arteries.

The possibility of using an exclusively percutaneous strategy to deliver foreign DNA to normal and balloon-dilated atherosclerotic arteries was studied by analysis of transfection efficiency in a rabbit model. A total of 22 external iliac arteries from 22 rabbits (10 normal and 12 atherosclerotic) were transfected with a solution of luciferase expression vector plasmid and liposome, using a dual balloon-catheter system. Analysis of the transfected segments revealed luciferase activity in 10 of the 22 arteries (4/10 normal vs 6/12 balloon-injured atherosclerotic, P = NS); no activity could be detected in the contralateral limb arterial segments used as controls. Luciferase activity levels in successfully transfected segments measured 4.10 +/- 1.19 (m +/- SEM) Turner light units (TLU), with 3.03 +/- 1.16 TLU found in normals vs 4.81 +/- 1.87 TLU in balloon-injured atherosclerotic arteries (P = NS). In situ hybridization of successfully transfected atherosclerotic sections showed expression of the luciferase gene mRNA from rare cells (less than 1/1,000) limited to the neointimal lesion. Thus, expression of new genetic material may be achieved in both normal and balloon-dilated atherosclerotic arteries following an exclusively percutaneous approach. The low efficiency of the current delivery strategy, however, represents a potential limitation that must be improved if this strategy is to be applied as a therapeutic approach to human vascular disease.


Interphotoreceptor matrix in the human retina: cone-like domains surround a small population of rod photoreceptors.

The interphotoreceptor matrix (IPM) in mammalian retinas is subdivided into rod and cone specific compartments: peanut agglutinin (PNA) binding glycoconjugates are associated with cones, whereas wheat germ agglutinin (WGA) binding glycoconjugates are associated with rods. To establish the identity of a photoreceptor cell type in the human retina with rod dimensions but with a matrix domain which stains with PNA, double label studies, using PNA-ferritin to decorate the extracellular domains and immunocytochemical techniques using a rod specific anti-opsin antibody were conducted. The PNA-binding domains were observed in the cone-associated IPM as well as in the IPM surrounding a small population of rod-shaped photoreceptors. The outer segments of these rod-shaped photoreceptors showed intense labeling with a rod specific anti-opsin antibody as did all other rods which were free of PNA-labeling. A quantitative analysis of all retinal quadrants indicates that this novel rod represents approximately 0.3% of the total rod population in the human retina.


TNF in combination with GM-CSF enhances the differentiation of neonatal cord blood stem cells into dendritic cells and macrophages.

We describe dendritic cell progenitors within the CD34+ stem cell compartment in neonatal cord blood and identify growth factors contributing to their differentiation. Granulocyte-macrophage colony-stimulating factor (GM-CSF), although mainly promoting the growth and differentiation of monocyte-macrophages (mono-m psi s), also induced the differentiation of cells with the distinctive morphological features of dendritic cells (DCs). Tumor necrosis factor (TNF) in combination with GM-CSF promoted further growth of both cell types but most notably increased the DC content. In situ analysis revealed that the cells exhibiting DC morphology were positive for class II major histocompatibility complex antigens but were CD14 negative, did not exhibit nonspecific esterase activity, and were nonphagocytic. Moreover, the mixed leukocyte reaction stimulatory capacity of cultures with the higher DC content was greater. TNF, interleukin-1 (IL-1), IL-6, or platelet-derived growth factor (PDGF) was inactive in promoting stem cell proliferation or DC morphology. IL-1 or PDGF synergized with GM-CSF to increase mono-m psi-associated cell proliferation but did not increase the DC content. The development of a common DC-monocyte precursor was suggested by the presence of colony-forming unit-like clusters containing mono-m psi s and DCs and one sharp proliferative peak. The loss of DC morphology after 21 days, coupled with increases in mono-m psi-associated markers and a constant number of viable cells, further suggests that DC morphology may fluctuate in culture or is a transient feature acquired by certain cells of the mono-m psi lineage.


High-frequency analysis of the signal-averaged ECG. Correlation with left ventricular mass in rabbits.

Standard electrocardiographic (ECG) criteria have exhibited poor correlation with left ventricular mass and poor sensitivity for left ventricular hypertrophy at acceptable levels of specificity. To assess the ability of the high-frequency filtered signal-averaged ECG to improve ECG correlation with left ventricular mass, signal-averaged orthogonal lead recordings in 29 normal rabbits and seven rabbits with left ventricular hypertrophy due to chronic aortic regurgitation were compared with left ventricular mass corrected for body weight. Voltage of the vector QRS complex was integrated over the total duration of the QRS after separate filtering with standard frequency (0-100 Hz) low-pass and high-frequency (44 Hz) high-pass filters. Measurement of individual X, Y, and Z lead R and S wave voltage was performed on averaged, standard frequency filtered complexes, and the maximal spatial vector magnitude was determined from the standard frequency filtered vectors. Voltage of the 44 Hz high-pass filtered vector QRS complex integrated over the total duration of the QRS (high-frequency vector integral) correlated closely with indexed left ventricular mass (r = 0.84, p less than 0.0001), significantly better than the correlation of standard frequency vector integral or maximal spatial vector magnitude voltages (r = 0.35 and r = 0.61, each p less than 0.01 vs high-frequency vector integral) and the correlation of orthogonal lead X R wave or lead Y S wave voltages (r = 0.55 and r = 0.37, respectively, each p less than 0.01 vs high-frequency vector integral).(ABSTRACT TRUNCATED AT 250 WORDS)


Influence of size of regions of interest on PET evaluation of caudate glucose consumption.

The aim of this study was to analyze the influence of variations in the size of regions of interest (ROIs) on values of caudate glucose consumption [regional cerebral metabolic rate of glucose (rCMRglc)] assessed by PET. In addition, we tried to establish the influence of ROI size on levels of significance assessing differences in mean caudate glucose consumption between two groups of subjects. For this purpose, rCMRglc was measured using [18F]fluorodeoxyglucose and the PC-4096 PET camera with an in-plane resolution of 7.1 mm in 12 normal subjects and 12 subjects with early Huntington disease. Caudate rCMRglc was histographically determined using 10 different ROI widths ranging from 2 mm, corresponding to the peak value of caudate rCM-Rglc, to 20 mm. The increase in ROI width from 2 to 20 mm led to a significant decrease of caudate rCMRglc by approximately 66% in the normal subjects and in the patients. The Student t value assessing the differences in mean caudate rCMRglc between the two groups decreased gradually from 5.61 for an ROI width of 2 mm to 3.78 for an ROI width of 20 mm. This corresponds to a worsening of the level of significance from 10(-5) to 10(-3), i.e., by a factor of 100. These data show that (given the resolution of presently used PET cameras) the selection of peak values of caudate rCMRglc is the best way to discriminate between groups of subjects supposed to differ with respect to caudate rCMRglc.


Segmental colonic dilatation in a neonate.

Segmental dilatation of the colon is rare, usually affecting older children, and may mimic Hirschsprung's disease. A neonate with Down's syndrome and malrotation presented with this condition and was cured by segmental resection.


Radiocontrast-induced nephropathy: current status and future prospects.

Radiocontrast-induced nephropathy (RCIN), a leading cause of in-hospital acute renal failure, is an acute decrease in renal function related to intravascular administration of iodinated radiocontrast agents. Though RCIN is relatively uncommon in patients without predisposing factors, patients with preexisting renal dysfunction, diabetes mellitus and severe congestive heart failure are at increased risk for acute renal failure following radiocontrast. Three recently developed animal models have provided important insights into the pathophysiology of RCIN. Specifically, these studies have implicated transient renal ischemia, direct renal tubular toxicity and changes in glomerular capillary permeability as possible mediators of RCIN, and these pathophysiologic mechanisms are not mutually exclusive. There is currently no effective treatment for RCIN. Assuring adequate hydration may reduce the risk of RCIN. In addition, synthetic atrial natriuretic factor and/or mannitol are promising, but as yet unproven, approaches to the prophylaxis of RCIN.


Control of drug release with a combination of prodrug and polymer matrix: antitumor activity and release profiles of 2',3'-diacyl-5-fluoro-2'-deoxyuridine from poly(3-hydroxybutyrate) microspheres.

Drug release was controlled by a combination of prodrug and polymer matrix. Prodrugs of 5-fluoro-2'-deoxyuridine with different physicochemical properties were synthesized by esterification with aliphatic acids (propionate, n-butyrate, and n-pentanoate). Microspheres containing these ester prodrugs were prepared with poly(3-hydroxybutyrate) of three molecular weights (65,000, 135,000, and 450,000). The release rates from the spheres depended on both the lipophilicity of the prodrug and the molecular weight of the polymer. Regardless of the polymer, the relative release rates were propionate greater than butyrate greater than pentanoate. The release of butyrate and pentanoate from the spheres consisting of low-molecular-weight polymer (M(r), 65,000) was faster than that from the spheres of higher molecular weight (M(r), 135,000 or 450,000). A single intraperitoneal injection of spheres of the highest molecular weight polymer containing butyrate or pentanoate resulted in higher antitumor effects against P388 leukemia in mice than did free prodrugs given over a period of five consecutive days. The polymer sphere itself showed low toxicity to and good biocompatibility with mice and rats.


Initial and delayed circulatory responses to orthostatic stress in normal humans and in subjects with orthostatic intolerance.

Gravitational stresses, which are common daily event for humans, result in a diminution in central blood volume, due to displacement of blood to the lower parts of the body. They demand complex adjustments in the cardiovascular system to offset the decrease in cardiac filling pressure. Such changes are necessary to sustain arterial blood pressure at an appropriate level so that there is adequate perfusion of vital organs, especially the brain. These adjustments must compensate for both the initial and sustained orthostatic stress. The rapid short-term adaptations are mediated primarily by the cardiovascular reflexes with humoral agents reinforcing these reflexes during severe and prolonged orthostatic stress. Understanding these complex reflex adjustments in normal humans is necessary in order to appreciate subjects with orthostatic intolerance.


Antigen presenting capacity of human decidua: no evidence for human decidual antigen presenting cell mediated immunoregulation.

Decidual antigen presenting cell (APC) mediated maternal immunoregulation has been reported. In the present study the ability of villous chorion as well as fetal cell pulsed early human pregnancy decidual APC to generate selectively antigen non-specific and MHC class II unrestricted CD8 positive T suppressor cells was reassessed in view of the fact that placental trophoblast, unlike the fetus, constitutes the fetal tissue of major contact at the maternal-fetal interface. Neither fetal cell nor villous chorion pulsed decidual APC generated maternal T cells with the ability to immunosuppress PHA-, Con A- and PWM-induced autologous or allogeneic lymphoproliferation. In only 2 out of 45 assays with villous chorion pulsed decidual APC was significant inhibition of mitogen induced lymphoproliferation detected and on no occasion with fetal cell pulsed decidual APC. No change in CD4/CD8 ratio of the maternal putative regulatory cells was detected by FACS analysis compared with control cultures. These findings suggest that decidual APC mediated immunoregulation plays no role in directing the maternal immune response.


Monoclonal antibodies identify Fc gamma receptors on unfertilized human oocytes but not spermatozoa.

Oolemmal Fc receptors have previously been shown to play a role in the promotion of adhesion by antibody labeled human spermatozoa to zona-free hamster eggs. In this work, we demonstrated the presence of Fc gamma RI, Fc gamma RII and Fc gamma RIII on the oolemma of unfertilized human oocytes by means of monoclonal antibodies directed against these receptors, detected both by immunobead rosetting and indirect immunofluorescence. These receptors were also functionally active in that they were able to bind human aggregated IgG, human IgG-Fc, mouse IgG1 and IgG2a. While the presence of oolemmal IgG-Fc receptors might play a role in reproductive failure, by their promotion of polyspermic fertilization, in cases where antisperm antibodies bound to the spermatozoan surface, their role in the normal physiology of fertilization or in other events unrelated to sperm incorporation remains to be determined. In contrast, Fc gamma receptors were not present on human spermatozoa, irrespective of their functional state (fresh ejaculated, capacitated or acrosome reacted).


Evidence for the expression of granulocyte-macrophage colony-stimulating factor receptors by human first trimester extravillous trophoblast and its response to this cytokine.

The present study has demonstrated that human extravillous trophoblast cells isolated from first trimester placentae can bind granulocyte-macrophage colony-stimulating factor (GM-CSF). We have used a technique which incorporates a phycoerythrin (PE)-conjugated GM-CSF in association with a monoclonal antibody against trophoblast (BC-1) in single and double flow cytometric analysis. Trophoblast cells are also observed to show increased DNA synthesis in response to GM-CSF as assayed by [3H]thymidine incorporation and proliferative activity as demonstrated by double immunocytochemical staining of cells with a trophoblast cytokeratin marker (PKK1) and for the nuclear proliferation antigen (Ki-67). These findings provide evidence that human extravillous trophoblast expresses receptors for GM-CSF and responds to this cytokine.


Enrichment of gamma delta T lymphocytes in human semen: relation between gamma delta T cell concentration and antisperm antibody status.

The presence and relative concentrations of T lymphocytes bearing the alpha beta and gamma delta cell receptor (TCR) in human semen were assessed and related to the occurrence of auto-antibodies to sperm in semen and peripheral blood. Using an immunoperoxidase technique, and monoclonal antibodies to the beta chain and delta chain of the human TCR, both alpha beta and gamma delta T cells were detected in each of 30 semen samples examined. In seven men with antisperm antibodies both on their ejaculated sperm and in their serum, the mean concentrations of gamma delta and alpha beta T cells were 3,560 and 3,230 cells/ml semen, respectively. In seven men with antisperm antibodies in serum only, the concentrations of gamma delta and alpha beta T cells were 860 and 1,280 cells/ml, while in 16 men with no evidence of auto-immunity to sperm there was a mean of 350 gamma delta T cells and 610 alpha beta T cells/ml. In contrast, the concentrations of gamma delta and alpha beta T cells in peripheral blood from these same men were unrelated to antisperm antibody status. The mean ratio of alpha beta to gamma delta T cells in peripheral blood of all subjects was 12. The ratio of alpha beta to gamma delta T cells in semen were 0.9 for men with sperm-bound and serum antisperm antibodies, 1.5 for men with antisperm antibodies in serum only and 1.7 for men lacking these auto-antibodies. These results were confirmed by FACS analysis. Thus, gamma delta T cells in human semen comprise a greatly increased proportion of the total T cell population as compared to the circulation. In addition, the relative and absolute concentration of gamma delta T cells are further elevated in semen from men with evidence of localized auto-immunity to their own sperm. These results suggest that gamma delta T cells may function in immune surveillance in the non-sterile proximal portions of the male genital tract and that replication of T cells bearing the gamma delta TCR accompanies an autoimmune response to sperm.


Freeze-fracture study of the site of attachment of Cryptosporidium muris in gastric glands.

The mode and organization of the attachment site of Cryptosporidium muris to gastric glands of stomach were investigated by the freeze-fracture method. Cryptosporidium muris was enveloped by a double membrane, of host plasma membrane origin, which formed the parasitophorous vacuole. The outer membrane of the double membrane was continuous with host plasma membrane, while the inner membrane was connected with the anterior part of the parasite plasma membrane at the annular ring. The density of intramembranous particles (IMP) was severely altered at the above two junctures. The parasitophorous outer membrane showed low IMP-density when compared to the host plasma membrane, although both membranes were continuous at the dense band. The inner membrane had few IMP, whereas the parasite plasma membrane showed numerous IMP, although both membranes were continuous at the annular ring. The size of dense band and annular ring was similar in diameter. The feeder organelle was clearly visible as membrane folds in freeze-fracture and some of them were connected with small vesicles of cytoplasm, indicating that the feeder organelle may play an important role for incorporation of nutrients from the host cell.


Pulmonary hypertension and cardiac insufficiency in three cows with primary lung disease.

Increased pulmonary arterial pressures as a result of pulmonary disease are described in two cows with chronic pneumonia and one cow with acute pneumonia. Based on clinical signs of congestive right heart failure, increased pulmonary arterial pressure, and right ventricular hypertrophy, cor pulmonale was diagnosed in one cow. Two cows had increased pulmonary arterial pressure and signs of right heart insufficiency, but right ventricular hypertrophy was not identified. Two of the cows had ventral edema and exercise intolerance. All cows had jugular venous distention and increased right atrial and pulmonary arterial pressures. Peripheral arterial PaO2 was decreased in two cows, and not measured in the third cow. Although an uncommon cause of congestive heart failure in cattle at low altitudes, pulmonary hypertension should be considered in cattle with clinical right heart failure.


Endoscopic treatment of endometriosis-associated infertility. Therapeutic, economic and social benefits.

In this report on the therapeutic results in patients with moderate and severe endometriosis treated by operative laser laparoscopy, we also present a comparative analysis of cost and duration of hospitalization and of convalescence in comparable patients treated by laparoscopy versus laparotomy. Sixty patients treated by the same surgeon with operative laser laparoscopy were followed for a period of at least one year to calculate cumulative pregnancy rates, monthly fecundity rates, and monthly probability of pregnancy. The mean duration of hospitalization and incapacitation as well as physician and hospital costs incurred by the laparoscopy treated patients were compared with those incurred by 60 patients with similar degrees of endometriosis but treated by microsurgery at laparotomy. In the laparoscopy group, 36 patients had stage III and 24, stage IV endometriosis. Monthly fecundity rates (6.7%), monthly probability of pregnancy (12.6%) and cumulative pregnancy rates (70.5%) did not differ between patients with stage III and IV disease. The total number of hospital days required by the 60 laparoscopy patients was 72 versus 258 for the laparotomy patients (P less than .001). The total number of days' incapacitation for laparoscopy patients was 216 versus 1,284 for the laparotomy group (P less than .001). The total cost of medical care was $223,260 for the laparoscopy group and $424,500 for the laparotomy group (P less than .001). Our results validate the therapeutic efficacy of operative laparoscopy in the treatment of moderate and severe endometriosis and confirm the substantial economic and social benefits of laparoscopy surgery over laparotomy.


Laparoscopic aspiration of ovarian endometriomas. Effect with postoperative gonadotropin releasing hormone agonist treatment.

In the period 1988-1990 this prospective study of 33 women with moderate or severe endometriosis who underwent laparoscopy for infertility and/or chronic pelvic pain, was conducted to evaluate the efficacy of aspirating endometriotic cysts followed by administration of a gonadotropin releasing hormone (GnRH) agonist in reducing the size of ovarian endometriomas. The cysts (mean diameter, 4.5 cm; range, 2-7; unilateral, 21 cases; bilateral, 12 cases) were punctured, aspirated, washed and emptied completely. After laparoscopy, 15 subjects received goserelin administered as a 28-day subcutaneous depot for three months, whereas 18 patients undergoing simple observation constituted internal controls. Ultrasound scans were performed before and at one, three and six months after laparoscopy. One case and three controls requested surgery between the four- and five-month follow-up scans and did not complete the study. All the other women had recurrent cysts at the six-month scan. There were no significant differences in mean endometrioma diameter between the two groups at any observation time nor between prelaparoscopic and six-month ultrasound examinations within each treatment group. We conclude that aspiration and washing of endometriotic cysts, combined with postoperative administration of GnRH agonists or not, is ineffective.


Laparoscopic appendectomy using a single umbilical puncture (minilaparoscopy).

The surgical outcome of the first 25 patients in whom the Pelosi single-puncture laparoscopic appendectomy technique was employed demonstrates the new approach as a safe, inexpensive and effective alternative to the currently used multiple-puncture method. The results suggest single-puncture (minilaparoscopy) operative endoscopy as the ultimate goal in the progression of minimally invasive surgery.


Management of ovarian masses. AAGL 1990 survey.

The American Association of Gynecologic Laparoscopists (AAGL) membership was surveyed on the use of laparoscopy in the management of persistent ovarian masses in 1990. A total of 13,739 laparoscopies were performed for this indication. Ninety-six percent of the respondents performed laparoscopy for this indication on premenopausal women only. Among respondents performing laparoscopy for suspected cancer, there was a 14% conversion rate to laparotomy, compared to 9% among those who performed direct laparotomies when cancer was suspected. An overall incidence of 4 per 1,000 cases of stage I ovarian cancer was found, and about 70% of women with persistent adnexal masses were managed by laparoscopy alone. The risks to women with cancer, as well as the benefits to those without, are discussed.


Management of adnexal masses by operative laparoscopy. Selection criteria.

The management of adnexal masses by operative laparoscopy is controversial. The application of strict criteria for preoperative patient selection and careful intraoperative assessment and management are critical to the appropriate use of this approach. Clinical examination, ultrasound imaging of the pelvis and the addition of the CA-125 tumor marker in postmenopausal women can aid in the selection of a population at low risk for malignancy that may be appropriate for operative laparoscopic adnexal surgery. Recommended procedures include careful intraoperative inspection of the pelvis and abdomen, liberal use of frozen sections, and conversion to immediate-staging laparotomy when malignancy is found.


Laparoscopic uterine suspension.

Laparoscopic uterine suspension is an effective method of correcting symptomatic uterine retroversion. Eighty patients with deep dyspareunia reproduced by palpation of their retroverted uterus underwent a modified Gilliam uterine suspension under laparoscopic visualization. There were no intraoperative or postoperative complications requiring laparotomy, and symptoms were relieved in 74 patients.


Incidence of bowel injury due to dense adhesions at the sight of direct trocar insertion.

A retrospective review of 4,532 outpatient laparoscopic procedures was conducted to determine the frequency of dense bowel adhesions at the level of the umbilicus. The umbilicus is the classical site of entry for the laparoscopic trocar. Using direct trocar insertion in all cases we encountered four bowel injuries. Bowel adhesion to the insertion site was noted in one case.


Jordan M. Phillips, M.D.--postdoctoral educator at large.

Twenty years ago Jordan Phillips recognized the importance of laparoscopy and aroused the interest of gynecologists by founding the American Association of Gynecologic Laparoscopists (AAGL). Within five years he had expanded the teaching mission of the AAGL to microsurgery, which revolutionized infertility surgery. Through his insatiable personal drive, he catalyzed the movement of people and ideas on an international level by organizing numerous postgraduate courses in North America, Europe and Asia, and later Australia and South America. Through his untiring efforts as an educator, Jordan Phillips has played a major role in preparing gynecologists in many parts of the world for the commanding position of advanced laparoscopic surgery, hysteroscopy and microsurgery in gynecologic practice.


Nd:YAG laser laparoscopic coagulation of symptomatic myomas.

The Nd:YAG laser dispersion effect, 2-5 mm in diameter, is utilized in a new laparoscopic procedure to coagulate and reduce symptomatic serosal and intramural myomas of moderate size (less than or equal to 10 cm). Depot leuprolide pretreatment for 2-6 months resulted in 40-60% shrinkage. Seventy-five patients 35-50 years old with symptomatic myomas, pain and pressure then underwent Nd:YAG laser coagulation for thorough devascularization of uterine myomas. Postoperative transvaginal ultrasound one, three and six months later showed the myomas reduced an average of 50-70% beyond the effect attributable to leuprolide. In two groups of patients whose myomas measured 5-10 cm and 3-5 cm after leuprolide pretreatment, laser coagulation subsequently reduced the myomas an average of 50%. In patients with postleuprolide myomas of 2-3 cm, virtually no myomas were identified postoperatively. The patients were followed up to 14 months. This laparoscopic procedure can be used in patients approaching menopause who wish to avoid abdominal myomectomy or hysterectomy.


Laparoscopic management of ovarian dermoid cysts.

Benign cystic teratoma (dermoid cyst) was managed laparoscopically in 25 cases (16 cyst excisions and 9 oophorectomies). Surgical procedures to avoid spill during ovarian cystectomy and oophorectomy were developed (14 cases). This series demonstrates a gradual evolution in surgical technique. Surgical outcome was good in all cases, complications were rare, and the procedure required a hospital stay less than 24 hours.


Direct binding of small nuclear ribonucleoprotein G to the Sm site of small nuclear RNA. Ultraviolet light cross-linking of protein G to the AAU stretch within the Sm site (AAUUUGUGG) of U1 small nuclear ribonucleoprotein reconstituted in vitro.

The major small nuclear ribonucleoproteins (snRNPs) U1, U2, U5 and U4/U6 participate in the splicing of pre-mRNA. U1, U2, U4 and U5 RNAs share a highly conserved sequence motif PuA(U)nGPu, termed the Sm site, which is normally flanked by two hairpin loops. The Sm site provides the major binding site for the group of common proteins, B', B, D1, D2, D3, E, F and G, which are shared by the spliceosomal snRNPs. We have investigated the ability of common snRNP proteins to recognize the Sm site of snRNA by using ultraviolet light-induced RNA-protein cross-linking within U1 snRNP particles. The U1 snRNP particles, reconstituted in vitro, contained U1 snRNA labelled with 32P. Cross-linking of protein to this U1 snRNA occurred only in the presence of the single-stranded stretch of snRNA that makes up the conserved Sm site. Characterization of the cross-linked protein by one and two-dimensional gel electrophoresis indicated that snRNP protein G had become cross-linked to the U1 snRNA. This was confirmed by specific immunoprecipitation of the cross-linked RNA-protein complex with an anti-G antiserum. The cross-link was located on the U1 snRNA by fingerprint analysis with RNases T1 and A; this demonstrated that the protein G has been cross-linked to the AAU stretch within the 5'-terminal half of the Sm site (AAUUUGUGG). These results suggest that the snRNP protein G may be involved in the direct recognition of the Sm site.


Refined 1.8 A crystal structure of the lambda repressor-operator complex.

The crystal structure of the lambda repressor-operator complex has been refined to an R-factor of 18.9% at 1.8 A resolution. This refinement, using data collected at low temperature, has revealed the structure of the N-terminal arm and shows that the interactions of repressor with the two halves of the pseudo-symmetric operator site are significantly different. The two halves of the complex are most similar near the outer edge of the operator site (in a region where the lambda and 434 repressors make similar contacts), but they become increasingly different toward the center of the operator. There are striking differences near the center of the site where it appears that the arm makes significant contacts to only one half of the DNA site. This suggested a new way of aligning the operator sites in phage lambda. The high resolution structure confirms many of the previously noted features of the complex, but also reveals a number of new protein-DNA contacts. It also gives a better view of the extensive H-bonding networks that couple contacts made by different residues and different regions of the protein, and reveals important new details about the helix-turn-helix (HTH) region, and the positions of many water molecules in the complex.


Effects of atrial natriuretic peptide on glycerol induced acute renal failure in the rat.

Acute and chronic experiments were performed in rats to examine whether atrial natriuretic peptide (ANP) has any beneficial effects on glycerol-induced acute renal failure (ARF). ANP infusion (Atriopeptin III, 1.0 microgram/kg+0.2 microgram/kg/min) improved the renal blood flow (RBF) and the glomerular filtration rate (GFR), and induced profound natriuresis in the early stage of ARF. By contrast, ANP decreased RBF in the control rats. In addition to these acute hemodynamic effects, long-term beneficial effects of ANP were also observed. A 75-min infusion of ANP significantly lessened the degree of azotemia as well as the extent of renal histologic damage assessed 24 hours after the glycerol injection. These results indicate that ANP can afford partial protection against both acute renal dysfunction and the chronic course of the glycerol-induced ARF, suggesting that ANP may be useful in the treatment of ARF.


Blood velocity patterns in poststenotic regions and velocity waveforms for myocardial inflow associated with coronary artery stenosis in dogs.

Velocity profiles across a vessel were investigated in poststenotic regions of the canine left coronary artery by our 80-channel 20 MHz ultrasound velocimeter. The velocity waveform in a small artery just before its penetration into myocardium was measured by our laser Doppler method. The poststenotic velocity configuration was characterized by a narrow region of high velocity with diastolic reverse flow near the wall which may dissipate energy. The velocity waveform in the distal small arteries exhibited increased systolic reverse flow with decreased diastolic forward flow, resulting in a remarkable reduction of coronary inflow into the myocardium.


Insights from in-vitro flow visualization into the mechanism of systolic anterior motion of the mitral valve in hypertrophic cardiomyopathy under steady flow conditions.

Hypertrophic obstructive cardiomyopathy is a heart disease characterized by a thickened interventricular septum which narrows the left ventricular outflow tract, and by systolic anterior motion (SAM) of the mitral valve which can contact the septum and create dynamic subaortic obstruction. The most common explanation for SAM has been the Venturi mechanism which postulates that septal hypertrophy, by narrowing the outflow tract, produces high velocities and thus low pressure between the mitral valve and the septum, causing the valve leaflets to move anteriorly. This hypothesis, however, fails to explain why SAM often begins early in systole, when outflow tract velocities are low or negligible or why it may occur in the absence of septal hypertrophy. The goal of this study was therefore to investigate an alternative hypothesis in which structural abnormalities of the papillary muscles act as a primary cause of SAM by altering valve restraint and thereby changing the geometry of the closed mitral apparatus and its relationship to the surrounding flow field. In order to test this hypothesis, an in vitro model of the left ventricle which included an explanted human mitral valve with intact chords and papillary muscle apparatus was constructed. Flow visualization was used to observe the ventricular flow field and the mitral valve geometry. Displacing the papillary muscles anteriorly and closer to each other, as observed clinically in patients with cardiomyopathy and obstruction produced SAM in the absence of septal hypertrophy. Flow could be seen impacting on the upstream (posterior) surface of the leaflets; such flow is capable of producing form drag forces which can initiate and maintain SAM.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of morphine on the phenotypic expression of cell surface markers on murine lymphocytes.

There is a growing literature indicating that opioid abuse by human addicts and opioid administration to animals have profound effects on the immune system. In the present study, implantation of morphine pellets in mice was associated with reduced phenotypic expression of the cell surface antigens specific to T-lymphocytes and to helper and cytotoxic/suppressor T-lymphocyte subtypes. The effect of morphine, as measured by flow cytometry using monoclonal antibodies specific for antigens expressed by these cells, was dose-dependent. The decrease in expression of antigens was apparent as early as 24 h after morphine pellet implantation and continued for 3 days. In addition, a time-dependent increase in the expression of these antigens was observed in placebo- and morphine-treated mice, suggesting that the pellets had a small antigenic effect. However, at all times studied, morphine-treated mice had fewer cells expressing the antigens than placebo-treated mice. Our results provide additional evidence that the use of opioids by IV drug abusers compromises their immune function.


Sequence comparison of the capsid region of hepatitis E viruses isolated from Myanmar and China.

Hepatitis E viruses (HEVs) were isolated during epidemics, one from Myanmar (formerly called Burma) and one from China and were partially sequenced. Another HEV Myanmar strain from sporadic hepatitis was previously sequenced by us. A cDNA sequence comparison was performed among them in the 3'-terminal region, approximately 750-base long. This region contained at least two immunological epitopes and was considered to correspond to the structural protein. The nucleotide sequence identity was 97.2% between the two Myanmar strains and 93.3 and 92.5% between the two Myanmar and the China strain. The deduced amino acid sequence identity ranged from 98.4 to 100.0% among the three strains. Thus this segment was well conserved on the amino acid level among the different strains isolated from these two Asian countries, although the China strain diverged more from the Myanmar strains on the nucleotide sequence level. This data may provide important information for the development of a vaccine and for identification of the virological link between different geographical locations.


Presynaptic serotonin mechanisms in rats subjected to inescapable shock.

After exposure to uncontrollable shock training, two distinct groups of rats can be defined in terms of their performance in learning to escape from a controllable stress. Learned helpless rats do not learn to terminate the controllable stress, whereas non-learned helpless rats learn this response as readily as naive control rats do. The present studies were designed to examine the correlations between the behavioral differences and the changes of presynaptic serotonergic activity, seen in these groups of rats. The major findings concerned presynaptic serotonergic effects in the hippocampus and hypothalamus of learned helpless rats. In the hippocampus, these included a statistically significant increase in three presynaptic 5-hydroxytryptamine (5-HT) mechanisms: K(+)-induced release of [3H]serotonin, high affinity uptake of [3H]serotonin and maximum density of binding sites for uptake of 5-HT, measured with [3H]paroxetine. In the hypothalamus, there was a differential modulation of all three presynaptic 5-HT mechanisms. A significant decrease in: K(+)-induced release of [3H]serotonin, in high affinity uptake of [3H]serotonin and the maximum binding site density of [3H]paroxetine binding was observed. No changes in uptake site binding was seen in other regions of the brain examined. These results implicate presynaptic serotonin mechanisms in the behavioral deficit caused by uncontrollable shock. In addition, a limbic-hypothalamic pathway may serve as a control center for the behavioral response to stress.


Mechanism of the anti-emetic activity of 5-HT3 receptor antagonists.

Ondansetron, a potent and highly selective 5-HT3 receptor antagonist, prevents emesis following chemotherapy by antagonising the action of 5-hydroxytryptamine (5-HT) at 5-HT3 receptors on vagal afferent neurons that innervate the gastrointestinal tract and 5-HT3 receptors in the central vomiting system. Evidence suggests that chemotherapy induces the release of 5-HT from enterochromaffin cells in the small intestine. This stimulates vagal afferent nerves via 5-HT3 receptors. Information is then relayed, via the vagus nerve, to the central vomiting system. 5-HT3 receptors are also found in the hind-brain vomiting system including the area postrema (the site of the chemoreceptor trigger zone for emesis). Therefore, following chemotherapy, 5-HT activates 5-HT3 receptors at 2 sites to induce emesis. Clinical data showing that a single dose of ondansetron prevents acute emesis suggest that it is important to block the initiation of the emetic reflex. This may prevent the recruitment of central mechanisms involving 5-HT3 receptors.


The effectiveness of a single intravenous dose of ondansetron.

This paper reviews data from 3 randomised, double-blind, parallel-group studies carried out in patients receiving high-dose cisplatin chemotherapy (50-120 mg/m2). These comparative trials show that a single intravenous dose of ondansetron (8-32 mg) is as effective as the continuous infusion and intermittent dose regimens used in previous clinical trials (8 mg i.v. followed by a 1 mg/h infusion for 24 h and 0.15 mg/kg i.v. x 3). One of the studies, carried out in Europe, demonstrated that a single 8 mg i.v. dose was as effective as 32 mg given either as an 8 mg loading dose followed by an infusion or as a single intravenous dose of 32 mg before chemotherapy. A similar study conducted in the United States showed that a 32 mg i.v. single dose was significantly more effective than both the 8 mg i.v. dose and the intermittent dose schedule. This study used a prospective stratification based on the dose of cisplatin (50-70 mg/m2 and greater than or equal to 100 mg/m2). In both strata the 32 mg dose was superior. These results emphasise the importance of selecting the dose of ondansetron (8-32 mg) based on factors that predispose patients to emesis, e.g., female gender, patients with a history of chemotherapy or motion sickness and the dose of cisplatin. The ondansetron dosing regimen for patients receiving a highly-emetogenic chemotherapy (8-32 mg i.v. followed by 8 mg orally twice daily) is both simple and flexible.


Ondansetron as prophylaxis for chemotherapy and radiotherapy-induced emesis in children.

Ondansetron was given as anti-emetic prophylaxis to 429 children receiving a variety of emetogenic cancer treatments for up to 8 days, in three, open, multicentre, European studies. Children aged between 6 months and 17 years with a variety of tumours and receiving chemotherapy or chemotherapy plus total body irradiation (TBI) were studied. Ondansetron was given intravenously, 5 mg/m2 or 8 mg, according to the surface area of the child, immediately before chemotherapy. Intravenous or oral treatment (2, 4 or 8 mg, according to surface area) was continued 3 times a day during chemotherapy or TBI, and for a further 2 days (non-cisplatin chemotherapy or TBI) or 5 days (cisplatin chemotherapy). The number of vomits and retches (each counting as an emetic episode) were recorded daily, as was an assessment of nausea, which was graded as none (not feeling sick at all), mild (feeling sick) or severe (feeling very sick). Responses were graded according to the number of emetic episodes during the worst 24-hour period. In addition, response was expressed in terms of emesis-free days as a proportion of all ondansetron treatment days. During chemotherapy, 66% of children experienced less than 3 emetic episodes on their 'worst day' and 88% had none or mild nausea. Sixty-eight percent of all ondansetron treatment days (2,131) were free of emesis. Of the patients who were poorly controlled with 'customary' anti-emetics, at least 81% experienced better control with ondansetron. When analysed according to the most emetogenic agent given 36, 59 and 75% of children reported less than 3 emetic episodes on their 'worst day' respectively, during cisplatin, ifosfamide and other less emetogenic chemotherapy. During conditioning for bone marrow transplantation with cyclophosphamide and TBI, 80 and 57% of patients, respectively, experienced less than 3 emetic episodes. The overall incidence of adverse events was low and headache (reported in 4% of patients) was the only event reported by more than 1% of patients. These studies show that ondansetron is a safe, well tolerated and an effective anti-emetic in the treatment of children receiving a wide variety of chemotherapy regimens.


Reduction of thyroid hormone receptor c-ERB A alpha mRNA levels in the hippocampus of Alzheimer as compared to Huntington brain.

A history of thyroid dysfunction has been cited as a possible risk factor for Alzheimer's disease (AD). Neurologic symptoms displayed by hypothyroid patients resemble, in part, those manifested by Alzheimer patients. To determine if a relationship exists between thyroid hormone receptor message levels and AD, in situ hybridization with tritiated antisense RNA probes for thyroid hormone receptors was used to examine the expression of these genes in Alzheimer and Huntington brain tissue. Message levels for a thyroid hormone receptor highly expressed in brain (c-ERB A alpha) was reduced by 52% in CA1 and 43% in CA2 in Alzheimer hippocampus as compared to Huntington controls. In contrast, message levels for another form of thyroid hormone receptor (c-ERB A beta 1) in Alzheimer hippocampus were not significantly different from Huntington controls. Temporal and cerebellar levels of c-ERB A alpha were elevated by 1.6-fold whereas temporal but not cerebellar levels of c-ERB A beta 1 were elevated 2.0-fold in Alzheimer brain. There was no correlation between thyroid hormone receptor levels and brain weight, autopsy interval, patient age, or the extent of neurofibrillary degeneration. Instead, decreased thyroid hormone receptor mRNA levels in Alzheimer-affected hippocampus were due to an increase in the percentage of neurons expressing lower message levels for these proteins.


Ondansetron compared with metoclopramide in the control of emesis and quality of life during repeated chemotherapy for breast cancer.

This was a multicentre, randomised, double-blind, parallel-group study which included female breast cancer patients, receiving their first of 6 scheduled courses of chemotherapy (cyclophosphamide greater than or equal to 500 mg/m2). Patients received an intravenous dose of 16 mg dexamethasone with either 8 mg ondansetron or 60 mg metoclopramide before chemotherapy, followed by oral dosing with 8 mg ondansetron or 20 mg metoclopramide 3 times daily for 5 days. A total of 93 patients were treated with ondansetron and 94 patients with metoclopramide. On day 1 of their first course of treatment 91 and 60% of patients in the ondansetron and metoclopramide groups respectively were free of emesis (p less than 0.001). Over the 5-day treatment period, the corresponding figures were 81 and 48% (p less than 0.001). The results for nausea also revealed highly statistically significant treatment differences (p less than 0.001) in favour of ondansetron for both day 1 and day 1-5 analyses of the first treatment course. Over the series of courses, 67% of patients receiving ondansetron completed all 6 courses with a maximum of 2 emetic episodes on their worst day, compared with 28% of patients receiving metoclopramide (p less than 0.001). A similar analysis for nausea revealed that 49% of patients receiving ondansetron completed all 6 courses with 'none' or 'mild' nausea compared with 27% of patients receiving metoclopramide (p less than 0.001). These differences were reflected in quality of life data (Rotterdam Symptom Checklist). After the first course of treatment, a statistically significant improvement (p = 0.002) in the psychological subscale scores was observed after ondansetron compared with metoclopramide. No differences were observed in the physical or functional activity subscales after the first course. However, the quality of life results over the series of courses revealed a more pronounced difference in favour of ondansetron in the psychological subscale scores (p less than 0.001) as well as trends in favour of ondansetron in the physical (p = 0.096) and functional activity (p = 0.056) subscales. Extrapyramidal symptoms were reported in 19% of patients in the metoclopramide group and resulted in 15% of patients withdrawing from their randomised anti-emetic schedule, either during or between treatment courses. Other adverse events were generally minor in nature and did not necessitate withdrawal from treatment. In conclusion, this study shows that ondansetron is significantly superior to metoclopramide (each with a single pre-treatment dose of dexamethasone) in the control of emesis over 6 courses of chemotherapy for breast cancer.(ABSTRACT TRUNCATED AT 400 WORDS)


A comparison of ondansetron with alizapride plus methylprednisolone in the control of cisplatin-induced emesis. The French Pneumology Group for the Ondansetron Study.

This randomised, single-blind, parallel-group study was carried out in 48 French pneumology centres to compare the anti-emetic efficacy of ondansetron and an alizapride plus methylprednisolone (ALI/MPS) combination in patients receiving high-dose cisplatin. A total of 220 patients were recruited of whom 209 were evaluable (100 on ondansetron and 109 on ALI/MPS). Thirty minutes before cisplatin, patients received either ondansetron (8 mg i.v.) or alizapride (4 mg/kg i.v.) combined with methylprednisolone (500 mg i.v.). The ondansetron and alizapride injections were repeated 4 and 8 h later. Thereafter, patients received oral ondansetron (8 mg) or alizapride (50 mg) 3 times daily for 5 days. Ondansetron was significantly superior to ALI/MPS in the control of acute emesis (p less than 0.001); 88/100 (88%) of ondansetron, and 69/109 (63%) of ALI/MPS patients experienced less than 3 emetic episodes. Similarly, ondansetron was superior to ALI/MPS for the control of acute nausea (visual analogue scale at 24 h; 13 vs. 22 mm respectively, p = 0.0012). The superiority of ondansetron over days 2-6 was not as great as that over the first 24 h, although there was a trend in favour of ondansetron. More patients treated with ondansetron wished to take the same anti-emetic treatment again (83% for ondansetron vs. 56% for ALI/MPS, p less than 0.001). Both treatments were well tolerated. This study shows that ondansetron is superior to a benzamide-corticosteroid combination in the control of acute cisplatin-induced emesis.


[Fixed drug-induced exanthema caused by novorin nose drops].

The case of a 36-year-old woman with typically fixed drug eruption is reported. A detailed case history and the diary of the patient helped to reveal the drug (Novorin nose drops) which caused the disease. It was proved by the lymphocyte transformation test.


The role of subendothelial laminin and platelet laminin receptors in haemostasis.

Laminin (LM) is a basement membrane glyco-protein which exhibits a number of biological activities, including the promotion of cell attachment and migration. In a static system, platelets attach to LM without spreading. In order to elucidate the mechanisms mediating platelet-LM interactions under flow conditions, we studied the effects of blocking subendothelial LM and its platelet receptor. We measured the extent of platelet adhesion to the endothelial cell extracellular matrix (ECM) using a parallel plate perfusion chamber. To do this, we performed the following experiments: i) blockade of subendothelial LM by incubation of ECM with an anti-LM antibody (Ab); ii) blockade of the platelet receptor for LM with: a) an anti-67 KDa receptor Ab; and b) three different LM-derived peptides (CFALRGDNP, IKVAV and CDPGYIGSR). In i) perfusates consisted of whole blood, whereas in ii) perfusates were prepared with washed platelets pre-incubated with Ab or peptides and reconstituted with plasma and washed red blood cells. Perfusions were carried out at 800 s-1 shear rate. Platelet deposition was morphometrically evaluated by a computerized system. Our results indicated that when ECM was pre-incubated with an anti-LM Ab (100 micrograms/ml, 30 min, 37 degrees C) a reduction of the surface coverage (SC) of 23.3 +/- 2.3% (p less than 0.01) was observed. When washed platelets were pre-incubated with an anti-67 KDa receptor Ab (40 micrograms/ml, 30 min, 37 degrees C) their interaction with ECM was decreased by 22.3 +/- 2.1% of SC (p less than 0.005). In other experiments platelets were pre-incubated with CFALRGDNP, IKVAV and CDPGYIGSR (200 micrograms/ml, 30 min, 37 degrees C).(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of adrenaline infusion on serum thromboxane B2 and plasma beta-thromboglobulin levels in hypertensive and normotensive subjects.

Adrenaline was infused in incremental doses of 0.05 up to 0.1 microgram/kg/min over a 60-min period in nine patients with mild essential hypertension and six age-matched normotensive controls. Blood samples were drawn at preset time intervals and plasma adrenaline, platelet count, serum thromboxane B2 (TxB2) and plasma beta-thromboglobulin (beta-TG) were measured. Adrenaline levels (m +/- SEM) rose significantly, from 0.078 +/- 0.01 (baseline) to 0.902 +/- 0.03 ng/ml (60 min), in the hypertensive group; a similar increase was observed in the control group (from 0.049 +/- 0.007 to 0.877 +/- 0.03 ng/ml). Platelet count increased significantly at early time points and remained high throughout infusion in both groups (hypertensive from 250 +/- 25 to 305 +/- 24 x 10(3)/microliters, control from 219 +/- 16 to 260 +/- 18 x 10(3)/microliters). TxB2 levels likewise increased significantly from 15 minutes after initiation of infusion. In hypertensive subjects the mean resting value of 186 +/- 17 ng/ml rose to 312 +/- 42 ng/ml, while in control subjects the resting value of 174 +/- 29 ng/ml rose to 286 +/- 32 ng/ml. Baseline levels of TxB2 were found to be higher in the hypertensive patients but not significantly. beta-TG levels increased from an initial value of 43.84 +/- 3.69 ng/ml to 59.5 +/- 4.69 ng/ml at 60 min in the hypertensive group, while a similar change from 28.7 +/- 19.2 ng/ml to 40.36 +/- 3.16 ng/ml was observed in the control group. These changes were significant, as was the difference between basal values in the two groups.(ABSTRACT TRUNCATED AT 250 WORDS)


5-Hydroxytryptamine1A receptor agonists in animal models of depression and anxiety.

The effects of different doses of buspirone, 3-dipropyl-amino-5-hydrochromar (NDO 008) and 8-hydroxydipropyl-aminotetralin (8-OH-DPAT) (administered intraperitoneally) were studied in tests of anxiolytic and antidepressant action in rats. These tests included the elavated plus maze test, the forced swim test, stress-induced suppression of open-field behavior, and the differential-reinforcement-of-low-rates-of-behaviour-72 sec (DRL 72 s) test. Buspirone (0.125 mg/kg) and NDO 008 (1.0 to 2.0 mg/kg) produced anxiolytic activity in the elevated plus maze, whereas 8-OH-DPAT did not in the doses employed. All three compounds increased activity in the forced swim test, although buspirone did so at a lower dose than NDO 008 and 8-OH-DPAT. In the stress-induced suppression test of open field activity all three compounds induced an antidepressant-like effect at different doses dependent on whether footshock (stressor) was presented 24 hr before or just prior to the open-field test. All three compounds even caused some reduction of activity in the non-shocked rats. 8-OH-DPAT (1.0 mg/kg) produced a significant and reliable increase in the Reinforcement/Response rate quotient in the DRL 72s test. These diverse results may provide an indication of potential clinical efficacy of the 5-HT1A agonists in the treatment of anxiety and depression.


[Primary amenorrhea revealing micropolycystic ovary syndrome].

Nine patients with polycystic ovary syndrome revealed by primary amenorrhea were studied clinically, biochemically and ultrasonographically. One of them had excess weight, four had hirsutism and five acne. Testosterone and delta 4-androstenedione levels were high in all patients. Luteinizing hormone (LH) basal levels were increased in five patients. Basal values of follicle-stimulating hormone (FSH) remained within normal range. The LH to FSH ratio was elevated (over 2) in six patients. The LSH response to gonadotropin-releasing hormone (GnRH) was explosive in all patients. Ultrasonography showed that three out of five patients had enlarged ovaries with multiple follicles not exceeding 8 mm. Following treatment, two patients had children after ovulation was induced with clomiphene citrate, one patient became pregnant spontaneously but thereafter opted for voluntary interruption; two patients are still on ovulation-inducing therapy; four patients still have contraception. These results indicate the clinical heterogeneity and the diagnostic problems that surround this uncommon form of a frequently observed disease.


[Digital necrosis in Raynaud's phenomenon revealing a testicular seminoma. Value of prostacyclin].

A testicular tumour could be diagnosed by the occurrence of a Raynaud's phenomenon complicated by severe digital arteritis. The arteritis rapidly regressed under prostacyclin therapy. Such vascular manifestations are frequent in testicular carcinoma, but they usually develop after chemotherapy. To our knowledge, this is the first case where they preceded the diagnosis and specific treatment of a tumour of the testis.


[Disorders of gastric emptying].

Disorders of gastric emptying are observed in many clinical situations. Their symptoms are diverse and correlate poorly with the objective abnormalities of gastric emptying. The underlying mechanism consists of abnormalities of basal electrical rhythm, fundic compliance, post-prandial antral motricity and, above all, antro-pyloro-duodenal co-ordination, associated to varying degrees. Among possible causes 3 clinical situations predominate: diabetes mellitus, functional gastrointestinal disorders (idiopathic dyspepsia) and sequelae of gastric surgery where retention of solids and accelerated evacuation of liquids may coexist in the same patient. Treatment of gastric incontinence rests, almost exclusively, on dietary measures, but several drugs, such as metoclopramide, domperidone and cisapride, are available to treat gastric stasis. Other compounds, notably motilin agonists (erythromycin and its derivatives) are currently being evaluated and will reinforce this therapeutic armentarium in a not too distant future.


[Electrocardiographic changes in patients with airway obstruction].

In severe bronchial asthma reversible electrocardiographic abnormalities are not rare. It is usually sinus tachycardia, right axis deviation, atrial enlargement and right bundle branch block. Transient ST-segment depression or elevation in inferior leads in severe acute asthma has been observed since long. Adrenergic stimulation, hyperventilation, hyperinflation and primary or secondary coronary insufficiency were as a causes. Severity of ECG signs correlated with the degree of airway obstruction. Our study was aimed at investigation of electrocardiographic abnormalities in chronic pulmonary obstructive disease and asthma and to assess the relationship of the extent of airway obstruction to the frequency of ECG changes. Correlation was found of ECG manifestation of sinus tachycardia, right ventricle hypertrophy. ventricular premature complex, right bundle branch block with the degree of airway obstruction.


[Surgical treatment of pneumothorax by video-thoracoscopy].

Between May and December 1991, 12 patients with spontaneous pneumothorax were treated surgically, using video-thoracoscopy. With this technique bullous lesions could be excised in 10 cases and pleurodesis could be performed in all patients. Morbidity and mortality were nil. The cosmetic and functional advantages of video-thoracoscopy were obvious. Long-term results remain to be evaluated. This technique has shown that it is possible in all cases to create pleurodesis (pleural poudrage or pleurectomy) and to treat parenchymatous lesions (excision of bullous systems). This suggests that the long-term results will be the same as those obtained with conventional surgery.


[Asymptomatic carriage of Chlamydia trachomatis in men consulting for condylomata acuminata].

A search for Chlamydia trachomatis by cell culture was carried out in the urethra of 82 male patients consulting for condyloma acuminata at the Clinical and Biological Centre for Sexually Transmissible Diseases of the Saint-Louis hospital, Paris. Three patients had discreet urethral signs, but none had urethral discharge. Cell culture was positive for C. trachomatis in 36 of the 82 patients (44 percent). This high prevalence suggests that C. trachomatis should systematically be looked for in the urethra of male patients consulting for condylomata acuminata. If this cannot be done, then a systematic treatment with tetracyclines should be instituted.


[Food habits of adults in France. Epidemiological data].

During the year 1990, 43,440 men and women aged from 17 to 69 years accepted to fill the NAQA questionnaire (18 items) on feeding habits, as part of a routine health examination. The frequency of consumptions and the amounts of food consumed are reported here according to sex. Mean energy intake was 2,586 kcal for men and 1,758 kcal for women and consisted of: fats 102 and 74 g respectively, sucrose 60 and 35 g, proteins 95 and 72 g, alcohol 24 and 4 g, calcium 1,128 and 982 mg and cholesterol 457 and 341 mg. For both men and women 16 percent and 16.8 percent respectively of non-alcohol intakes were made of proteins and 38 percent and 38.6 percent of lipids. Calcium intake was low in 7.5 percent of the subjects. Data analysis by sex and 10-year age groups showed a decrease of fats and sucrose rations and stability of protein rations as the subjects were growing older. These results were similar to those of a recent survey performed with the food history technique. The data obtained by us are an interesting source of information for Public Health epidemiology. They are compared with those found in the literature and discussed.


[Incidence of nosocomial infections in a military hospital].

Over a 2-year period, 479 cases of nosocomial infections were identified in our hospital by a surveillance method based on the bacteriological laboratory results. The monthly incidence rate ranged from 1.8 to 4 percent of all in-patients. With this method, a 2 percent background noise and a 4 percent alarm threshold could be defined. The most frequent nosocomial infections were urinary tract infections (77 percent), purulent skin infections (12 percent) and septicaemias (10 percent). Compared during 15 days with the most sensitive surveillance method of the National Nosocomial Infections Surveillance System, our method proved insufficient to detect nosocomial lung infections and superficial surgical wound infections. On the other hand, it was highly satisfactory to watch for urinary tract infections and septicaemias. Improvements that would not put a heavy burden on the work of clinical departments are suggested.


[Chylous ascites caused by retroperitoneal neoplastic obstruction of lymphatic vessels. Treatment by peritoneo-jugular shunt. A case].

A case of chylous ascites due to retroperitoneal tumoral compression is reported. Following failure of a medical treatment which consisted of paracentesis and medium-chain triglyceride (MCT) diet, the ascites dried up after installation of a peritoneum-jugular vein shunt valve. This operation is seldom performed in patients with chylous ascites since the medical treatment with paracenteses, MCT diet or even total parenteral nutrition succeeds in drying the effusion in almost 50 percent of the cases, notably those with postoperative lesion.


[Isolated atrial fibrillation. The risk of embolism and its prevention].

Whether or not atrial fibrillation is alone, if not idiopathic, is difficult to determine. The risk of embolization in lone atrial fibrillation is distinctly higher in healthy subjects over 60 years of age when the left atrium is dilated. In chronic atrial fibrillation this risk is higher than in paroxysmal fibrillation, especially within the year following the onset of the arrhythmia. In most patients anticoagulant therapy is effective in the primary or secondary prevention of embolic accidents. In subjects older than 75 aspirin given in daily doses of 325 mg seems to give similar results. The risk of antithrombosis therapy must not be underevaluated. The alternative is to maintain or restore the sinus rhythm, even at an advanced age, if the arrhythmia is recent and the left atrium is moderately dilated.


[Pregnant seropositive drug addicts].

Understanding actual experiences of sexuality and pregnancy in HIV positive drug addicts is essential for the prevention of AIDS dissemination. A rigorous organization and a coherent therapeutic project are needed for pregnancy supervision (pregnancy being carried on in over 50 percent of cases), for psychological preparation to the recognition of foetus reality and birth, and for newborn follow up to prevent his abandonment. The specific problems of pregnancy, seropositivity and drug addiction should be taken into account at the same time. The intervention of a voluntary, motivated and polyvalent team and a hospital opening towards outreach workers are two essential conditions of success.